Synergistic impact of -linolenic acid and -tocopherol on in vitro maturation and culture of buffalo oocytes

Abstract The objective of the current study was to investigate the synergistic impact of -Tocopherol and -Linolenic acid (100 µM) on IVM and IVC of Nili Ravi buffalo oocytes. Oocytes were obtained from the ovaries of slaughtered buffaloes within two hours after slaughter and brought to laboratory. Buffalo cumulus oocyte complexes were placed randomly in the five experimental groups included; GROUP 1: Maturation media (MM) + 100 µM ALA (control), GROUP 2: MM + 100 µM ALA + 50M -Tocopherol, GROUP 3: MM + 100 µM ALA + 100M -Tocopherol, GROUP 4: MM + 100 µM ALA + 200 M -Tocopherol and GROUP 5: MM + 100 µM ALA + 300 M -Tocopherol under an atmosphere of 5% CO2 in air at 38.5 °C for 22-24 h. Cumulus expansion and nuclear maturation status was determined (Experiment 1). In experiment 2, oocytes were matured as in experiment 1. The matured oocytes were then fertilized in Tyrodes Albumin Lactate Pyruvate (TALP) medium for about 20 h and cultured in synthetic oviductal fluid (SOF) medium to determine effect of -Linolenic acid (100 µM) and -Tocopherol in IVM medium on IVC of presumptive zygotes. To study the effect of -Linolenic acid (100 µM) in IVM media and increasing concentration of -tocopherol in the culture media on early embryo development (Experiment 3), the presumptive zygotes were randomly distributed into the five experimental groups with increasing concentration of -tocopherol in culture media. Higher percentage of MII stage oocytes in experiment 1(65.2±2.0), embryos at morula stage in experiment 2 (30.4±1.5) and experiment 3 (22.2±2.0) were obtained. However, overall results for cumulus cell expansion, maturation of oocyte to MII stage and subsequent embryo development among treatments remain statistically similar (P > 0.05). Supplementation of -tocopherol in maturation media having -Linolenic acid and/or in embryo culture media did not further enhance in vitro maturation of oocyte or embryo production.

Resumo O objetivo do presente estudo foi investigar o impacto sinérgico do -tocoferol e do ácido -linolênico (100 µM) na MIV e CIV de oócitos de búfala Nili Ravi. Os oócitos foram obtidos dos ovários de búfalos abatidos duas horas após o abate e levados ao laboratório. Complexos de oócitos cumulus de búfalo foram colocados aleatoriamente nos cinco grupos experimentais incluídos; GRUPO 1: Meio de maturação (MM) + 100 µM ALA (controle), GRUPO 2: MM + 100 µM ALA + 50 µM -tocoferol, GRUPO 3: MM + 100 µM ALA + 100 µM -tocoferol, GRUPO 4: MM + 100 µM ALA + 200 M -tocoferol e GRUPO 5: MM + 100 µM ALA + 300 M -tocoferol sob uma atmosfera de 5% de CO2 em ar a 38,5 °C por 22-24 h. A expansão cumulus e o estado de maturação nuclear foram determinados (Experimento 1). No experimento 2, os oócitos foram maturados como no experimento 1. Os oócitos maturados foram então fertilizados em meio de Tyrode's Albumina Lactato Piruvato (TALP) por cerca de 20 h e cultivados em meio de fluido oviductal sintético (SOF) para determinar o efeito do ácido -linolênico (100 µM) e -tocoferol em meio IVM em IVC de presumíveis zigotos. Para estudar o efeito do ácido -linolênico (100 µM) em meio IVM e aumentar a concentração de -tocoferol no meio de cultura no desenvolvimento inicial do embrião (Experimento 3), os presumíveis zigotos foram distribuídos aleatoriamente nos cinco grupos experimentais com concentração crescente de -tocoferol em meios de cultura. Maior porcentagem de oócitos em estágio MII no experimento 1 (65,2 ± 2,0), embriões em estágio de mórula no experimento 2 (30,4 ± 1,5) e experimento 3 (22,2 ± 2,0) foram obtidos. No entanto, os resultados gerais para a expansão das células do cumulus, maturação do oócito para o estágio MII e desenvolvimento embrionário subsequente entre os tratamentos permanecem estatisticamente semelhantes (P> 0,05). A suplementação de -tocoferol em meios de maturação com ácido -linolênico e / ou em meios de cultura de embriões não aumentou ainda mais a maturação in vitro de oócitos ou a produção de embriões.

Prevalence of gastrointestinal parasitic infection in cows and buffaloes in Lower Dir, Khyber Pakhtunkhwa, Pakistan

Abstract Gastrointestinal (GI) Parasitic infection is a hot issue for cattle management. There is variation of GI parasites effects in sex, age of cattle, drinking water condition, nutrition, and severity of infection. Studies on prevalence of GI parasites among cattle population in Dir Lower are lacking. A total of 40 farms were selected randomly in six tehsil namely Tehsil Adenzai, Tehsil Timergara, Tehsil Balambat, Tehsil Munda ,Tehsil Lalqala, Tehsil Khall. Freshly cattle fecal samples were collected randomly from the selected farms during March 2018 till December 2018. Out of 314 buffaloes and cattle examined 58.59% (184/314) were positive for eggs, cyst/oocyst of one or more species of GI Parasites. The prevalence of parasitic infection was higher in Buffaloes 63.55% (75/118) as compared to Cow 55.61% (109/196) but the difference was not significant (p>0.05) Entamoeba,spp, Moniezia spp, Haemonchus spp and Coccidian spp were found in this study. The non-treated animals indicated the highest percentage of infection in cow 57.71% (101/175) and buffalo 68.13% (62/91).GI parasite prevalence in female animal were higher female cow 62.58% (87/139) and female buffalo 77.33% (58/75) as compared to male. But the difference is non-significant (p> 0.05) Yearling calves had the lower rate of GI parasitic infection than adults. Future investigations are necessary to evaluate the economic loss due to GI parasites in cattles.

Resumo A infecção gastrointestinal (GI) parasitária é um assunto importante para o manejo do gado. Existem variações nos efeitos dos parasitas GI quanto a sexo, idade do gado, condição da água potável, nutrição e gravidade da infecção. Faltam estudos sobre a prevalência de parasitas GI entre a população de gado em Lower Dir. Um total de 40 fazendas foi selecionado aleatoriamente em seis tehsil, nomeadamente Tehsil Adenzai, Tehsil Timergara, Tehsil Balambat, Tehsil Munda, Tehsil Lalqala, Tehsil Khall. Amostras fecais de gado fresco foram coletadas aleatoriamente das fazendas selecionadas de março de 2018 até dezembro de 2018. Dos 314 búfalos e bovinos examinados, 58,59% (184/314) foram positivos para ovos, cisto/oocisto de uma ou mais espécies de parasitas GI. A prevalência de infecção parasitária foi maior em Buffaloes 63,55% (75/118) em comparação com vaca 55,61% (109/196), mas a diferença não foi significativa (p > 0,05). Entamoeba spp, Moniezia spp, Haemonchus spp e Coccidian spp foram encontrados neste estudo. Os animais não tratados indicaram a maior porcentagem de infecção em vacas 57,71% (101/175) e búfalos 68,13% (62/91). A prevalência do parasita GI em fêmeas foi maior em vacas fêmeas 62,58% (87/139) e búfalas 77,33% (58/75) em comparação ao masculino. Mas a diferença não é significativa (p > 0,05). A modelagem linear geral mostrou que o tratamento do animal estava significativamente relacionado com a prevalência de parasitas GI. Bezerros de um ano tiveram a maior taxa de infecção parasitária gastrointestinal. Futuras investigações são necessárias para avaliar a perda econômica devido aos parasitas GI em bovinos.

Effect of incorporation of aloe vera (Aloe barbadensis Miller) on the physicochemical and bromatological characteristics and shelf-life of buffalo milk yogurt / Efecto de la incorporación de Aloe vera (Aloe barbadensis Miller) en las características fisicoquímicas, bromatológicas y vida útil de un yogurt de leche de búfala

ABSTRACT In the dairy industry, fermented products are traditionally made with cow's milk, however, recently other ingredients have been used that give different qualities to the final product; among them is buffalo milk, which has distinctive characteristics and is rarely utilized industrially in Colombia, for this reason, the present study aimed to evaluate the effect of adding aloe vera (Aloe barbadensis Miller) on the physicochemical and bromatological properties and shelf-life of buffalo milk yogurt. First, an aloe vera sauce was made, which was added to buffalo milk yogurt in three percentages (5, 10, and 15 %), and one was chosen using a preference ordering test. Subsequently, the selected product was evaluated for physicochemical and bromatological parameters and mineral content. The shelf-life was also calculated using an accelerated shelf-life study at temperatures of 5, 15, 25, and 35 °C. According to the sensory panel the best yogurt was the sample with 15 % of aloe vera sauce. For the bromatological analysis, it was found that the final product had 2.57 % protein, 5.87 % fat, 0.49 % ash, and 74.9 % moisture. Shelf-life calculations yielded an average of 28.6 days. It is concluded that aloe vera positively impacted the bromatological parameters and mineral content of buffalo milk yogurt.

RESUMEN En la industria láctea, se elaboran productos fermentados, tradicionalmente, con leche de vaca; sin embargo, en los últimos tiempos, se han utilizado otros ingredientes, que dan diferentes cualidades al producto final; entre ellos, se encuentra la leche de búfala, que tiene características únicas y además es poco utilizada industrialmente en Colombia. La presente investigación tuvo por objetivo evaluar el efecto de la adición de aloe vera (Aloe barbadensis Miller), en las características fisicoquímicas, bromatológicas y vida útil de un yogurt de leche de búfala. Primeramente, se realizó una salsa de aloe vera, la que se le agregó al yogurt de leche de búfala, en tres porcentajes (5, 10 y 15 %) y se escogió uno, mediante una prueba de preferencia por ordenación. Posteriormente, al producto seleccionado, se le evaluaron los parámetros fisicoquímicos, bromatológicos y contenido de minerales. También fue calculada la vida útil del yogurt, utilizando un estudio de vida útil, acelerado a temperaturas de 5, 15, 25, y 35 °C. De acuerdo con el panel sensorial, se estableció que la mejor muestra de yogurt fue aquella que tenía 15 % de salsa de aloe vera. En cuanto al análisis bromatológico, se encontró que el producto final tuvo 2,57 % de proteína, 5,87 % de grasa, 0,49 % de cenizas y 74,9 % de humedad. Los cálculos de vida útil arrojaron 28,6 días en promedio. Se concluye que el aloe vera impactó positivamente los parámetros bromatológicos y el contenido de minerales del yogurt de leche de búfala.

Application of lectins for detection of glycomic differences in the epididymal duct in water buffalo (Bubalus bubalis) and dromedary camel (Camelus dromedarius) / Aplicación de lectinas para la detección de diferencias glucómicas en el conducto epididimario en búfalos de agua (Bubalus bubalis) y dromedarios (Camelus dromedarius)

SUMMARY: The present study was conducted to detect the differences in glycohistochemical features in the epididymal duct of the dromedary camel and the water buffalo. Epididymal sections (caput, corpus and cauda) from both species were stained with fluorescein isothiocyanate (FITC) conjugated lectins. Binding sites for five lectins (DBA, GSA-1, HPA, PNA and WGA) have been found in both species. The binding sites of different lectins showed significant variations in the pattern of distribution in both a species. This included both species-specific and region-specific order. Additionally, only three (GSA-1, PNA and WGA) out the five lectins studied exhibited binding sites in all epididymal regions in both species. The other two lectins (DBA and HPA) followed the same order recorded for the other three (GSA-1, PNA and WGA) in buffalo, but failed to show any binding sites in cauda epididymis in camel. In conclusion, the variable regional and species-specific distribution features of lectins revealed that both species have diverse glycomic characteristics that may be related to their different reproductive patterns. However, the glycome-associated functional capacities remain obscured and need further profound investigations.

RESUMEN: El presente estudio se realizó para detectar las diferencias en las características glicohistoquímicas del conducto epididimal del dromedario y el búfalo de agua. Las secciones del epidídimo (cabeza, cuerpo y cola) de ambas especies se tiñeron con lectinas conjugadas con isotiocianato de fluoresceína (FITC). Se encontraron sitios de unión para cinco lectinas (DBA, GSA-1, HPA, PNA y WGA) en ambas especies. Los sitios de unión de diferentes lectinas mostraron variaciones significativas en el patrón de distribución en ambas especies. Esto incluía tanto el orden específico de la especie como el específico de la región. Además, solo tres (GSA-1, PNA y WGA) de las cinco lectinas estudiadas exhibieron sitios de unión en todas las regiones del epidídimo en ambas especies. Las otras dos lectinas (DBA y HPA) siguieron el mismo orden registrado para las tres restantes (GSA-1, PNA y WGA) en búfalos, pero no mostraron ningún sitio de union en la cola del epidídimo en camellos. En conclusión, las características de distribución regionales y específicas de especies variables de las lectinas revelaron que ambas especies tienen características glucómicas diversas que pueden estar relacionadas con sus diferentes patrones reproductivos. Sin embargo, las capacidades funcionales asociadas con el glicoma permanecen desconocidas y requieren mayor investigación.

Prevalence of Reproductive Disorders in Dairy Buffaloes of Sudoor Paschim Province (SPP), Nepal

Buffalo (Bubalus bubalis), also called black gold is an important species of husbandry animal in Nepal. Its multi utility roles in agriculture economy goes beyond providing milk, meat, manure, and draught power in many developing countries including Nepal. Buffalo faming is picking up commercial trend but issues, increasing cost of inputs, increasing resistance and troubleshooting of infective agents remains major hurdles to economic returns. A cross-sectional study using structured questionnaire survey was conducted to determine the incidence of reproductive disorders in dairy buffaloes and its associated risk factors in Sudoor Paschim Province (SPP) of Nepal from November 2020 to February 2021. Out of 389 dairy buffaloes under investigation, 220 (56.56%) had encountered at least one of the reproductive disorders. The major reproductive disorders reported in the present study included repeat breeding (24.94%), followed by cervico-vaginal prolapse (8.23%), retention of placenta (6.68%), uterine prolapse (5.40%), dystocia (4.63%), abortion (2.83%), uterine torsion (2.57%) and still birth (1.29%). Overall, it is suggested that improvement in management system, breeding system, accurate heat detection, balanced feeding, and hygienic condition should be done to minimize the reproductive health disorders in buffaloes of Sudoor Paschim Province

Expression Assessment of Some Immunity-related Genes in Buffalo Infected with Endometritis

Background and Aim: Despite the economic importance of buffalo as a main source of milk and meat, only little attention has been directed to its immune and reproductive performance. The early diagnosis of subclinical endometritis may reduce the economic loss of buffalo’s production. The difference in expression profiles of immunity-related genes has an important role in the early detection of subclinical endometritis. This study aimed to assess the expression of five immunity-related genes: TGFBR1, PTGER2, PTGER4, HP and CXCL5 in endometritis-infected buffaloes. Materials and Methods: Total RNA was extracted from 120 buffalo uterine samples; 60 infected with endometritis and 60 healthy ones. Qt-PCR was performed on cDNA synthesized from extracted RNA using Sybr green and GAPDH as a house-keeping gene. Results: The results showed the up-regulation of two tested genes; TGFBR1 and CXCL5 in endometritis-infected buffalo compared to healthy animals by 7.9 and 4.3 folds, respectively at a significance level of p<0.05. The other three tested genes; PTGER2, PTGER4 and HP were down-regulated in buffalo during endometritis infection at different levels; PTGER2 and HP (0.6 folds, p<0.05) and PTGER4 (0.4 fold, p= 0.2). Conclusions: It is to be concluded that the assessment of expression of inflammation-related immunity genes may have an effective role on the detection of endometritis infection in buffalo during its early stages and this early diagnosis can reduce the economic loss of buffalo production and reproduction.

Desenvolvimento de bebidas lácteas probióticas adicionadas de polpa de morango e suplementadas com L-triptofano elaboradas a partir de leite bubalino e bovino / Development of probiotic dairy beverages with strawberry pulp and supplemented with L-tryptophan from buffalo and bovine milk

O objetivo deste estudo foi desenvolver uma formulação de bebida láctea bubalina probiótica adicionada de polpa de morango, comparando os efeitos do uso do leite de búfala e de vaca na elaboração dos produtos e verificando a possibilidade de suplementação com triptofano nos produtos lácteos probióticos. Como primeira etapa do trabalho, bebidas lácteas probióticas foram elaboradas a partir de leite bubalino e bovino, fermentadas com Streptococcus thermophilus TA040, Lactobacillus bulgaricus LB340 e Lactobacillus acidophilus La5, e formuladas com 0, 25 e 50% de soro em sua formulação. As bebidas foram avaliadas quanto à cinética de fermentação das culturas láticas utilizadas, ao teor de proteína, gordura e sólidos totais não gordurosos, pós-acidificação, viabilidade das culturas fermentadoras e sua capacidade de sobrevivência ao estresse gastrointestinal in vitro. As bebidas lácteas bubalinas apresentaram resultados superiores as bebidas bovinas. O uso do leite de búfala na elaboração das bebidas lácteas promoveu benefícios quanto as culturas láticas presentes nos produtos, exercendo efeito protetivo e influindo na preservação da viabilidade das bactérias ao longo do armazenamento refrigerado e durante a simulação do estresse gastrointestinal in vitro. As bebidas lácteas elaboradas com 25% apresentaram os resultados mais próximos aos obtidos pelos produtos controle, sem adição de soro, sendo selecionadas para a segunda parte do estudo. Nesta etapa, as formulações de bebida láctea com 25% de soro, foram acrescidas de um preparado com polpa de morango e bebidas sem adição da fruta, utilizadas como controle. As bebidas lácteas bubalinas frutadas, apresentaram menor teor de gordura e melhores características reológicas, com maior viscosidade e consistência do que os produtos controle, sem afetar a pós-acidificação, o perfil de ácido graxo, assim como, a viabilidade e a resistência às condições de estresse gastrointestinal in vitro das culturas fermentadoras. A avaliação da possibilidade de suplementar lácteos probióticos com triptofano foi realizada em conjunto com a Universidade de Milão. Para isso, iogurtes probióticos receberam adição de triptofano antes ou após a fermentação, sendo avaliados com relação ao perfil de pós-acidificação, quantidade de triptofano nos produtos, número de células viáveis por plaqueamento e citometria de fluxo ao longo do armazenamento a 25° e 4°C. Complementarmente, a influência da presença do triptofano no crescimento e produção de compostos antimicrobianos pelas culturas láticas, também foi avaliada. A adição de triptofano após a fermentação dos iogurtes, que foram armazenados sob refrigeração (4°C), além de não afetar a pós-acidificação dos produtos, apresentou benefícios quanto a viabilidade L. acidophilus, redução do dano e aumento do número de células vivas, promovendo teor maior do aminoácido nos iogurtes. A presença do triptofano nos meios de cultivo, também influenciou de forma positiva o crescimento de S. thermophilus e L. acidophilus, melhorando o desenvolvimento das bactérias durante a fermentação e influindo em uma maior atividade antilistérica por parte do S. thermophilus. Diante da influência positiva da aplicação do leite de búfala na elaboração das bebidas lácteas, assim como, a adição do triptofano em iogurtes probióticos, a suplementação do aminoácido em bebidas lácteas bubalinas frutadas permitiria a obtenção de um produto funcional, onde seus benefícios estariam relacionados tanto ao consumo do probiótico presente no produto quanto a complementação de triptofano na dieta do consumidor

The aim of this study was to develop a formulation of probiotic buffalo dairy beverage added with strawberry pulp, comparing the effects of using buffalo and cow's milk in the preparation of products and verifying the possibility of tryptophan supplementation in probiotic dairy products. As a first stage of the work, probiotic dairy beverages were made from buffalo and bovine milk, fermented with Streptococcus thermophiles TA040, Lactobacillus bulgaricus LB340 and Lactobacillus acidophilus La5, and formulated with 0, 25 and 50% whey in their formulation. The beverages were evaluated for the fermentation kinetics of the used lactic cultures, the levels of protein, fat and total no fat solids, post-acidification, fermenting cultures viability and their ability to survive gastrointestinal stress in vitro. Buffalo milk use in dairy beverages production promoted benefits regarding the lactic cultures present in the products, exerting a protective effect and influencing the viability preservation of bacteria during the cold storage and simulation of gastrointestinal stress in vitro. Dairy beverages made with 25% whey addition showed results similar to those obtained by the control products, without whey addition, being selected for the second part of the study. In this part, the dairy beverages formulations with 25% whey, were added with a preparation were added with a strawberry pulp preparation and dairy beverages without added fruit, used as a control. Fruity bubaline dairy beverages had lower fat content and better rheological characteristics, with higher viscosity and consistency than control products, without affecting post-acidification, fatty acid profile, as well as viability and resistance to in vitro gastrointestinal condition of fermented cultures. The possibility of supplementing probiotic dairy products with tryptophan was evaluated in partnership with the University of Milan. For this, probiotic yogurts received the addition of tryptophan before or after fermentation, being evaluated in relation to the post-acidification profile, tryptophan amount in the products, viable cell number per plating and flow cytometry during storage at 25°C and 4°C. In addition, the influence of the tryptophan presence on the growth and production of antimicrobial compounds by lactic cultures was also evaluated. The addition of tryptophan after the yogurt fermentation, which were stored under refrigeration (4°C), in addition to not affecting the post-acidification of the products, showed benefits to the viability of L. acidophilus, reduced the damage and increased the number of cells promoting higher amino acid content in yogurts. Tryptophan presence in the culture media also positively influenced the growth of S. thermophiles and L. acidophilus, improving the development of bacteria during fermentation and influencing better antilisteric activity in the part of S. thermophiles. In view of the buffalo milk positive influence observed after the application in dairy beverage preparation, as well as the addition of tryptophan in probiotic yoghurts, amino acid supplementation in fruity buffalo dairy beverages would allow to obtain a functional product, where its benefits would be related both to the consumption of the probiotic present in the product as to the supplementation of tryptophan in the consumer's diet

La suplementación estratégica de bucerros manejados en pasturas con predominio de Hymenachne amplexicaulis estimula las conductas de ingestión y mejora la ganancia de peso / Strategic supplementation of buffalo calves managed in pastures with a predominance of Hymenachne amplexicaulis stimulates ingestion behavior and improves weight gain

Introducción: En Colombia, la producción bufalina viene creciendo de forma importante. Los búfalos se adaptan bien a zonas aluviales, que se caracterizan por la presencia de especies de gramíneas nativas, como Hymenachne amplexicaulis. Objetivo: Evaluar el efecto de diferentes tipos de suplementación sobre el desempeño productivo y comportamiento ingestivo de bucerros manejados en pasturas con predominio de Hymenachne amplexicaulis. Materiales y métodos: Treinta bucerros Murrah, con 67.5 (DE=2) kg de peso y 4 meses de edad, se asignaron, aleatoriamente, sí son tres tratamientos: SS: sin suplementación; SM: suplemento mineral y SM+C: suplemento mineral + balanceado (0.5% peso corporal). Los animales pastorearon en praderas con predominio de Hymenachne amplexicaulis, y en la noche recibieron el suplemento en corrales. Las variables estudiadas fueron la ganancia de peso y el tiempo destinado a realizar diferentes conductas de ingestión, además de las tasas de bocados y estaciones de alimentación. Resultados: El consumo diario de mezcla mineral fue de 33.8 g en los dos grupos suplementados, y el de balanceado de 149.3 g. La oferta de SM y de SM+C produjo un incremento extra en la ganancia de peso de 68 y de 227 g, respectivamente (p <0.001). Los animales suplementados presentaron mayor tiempo de pastoreo (p<0.001) y menor tiempo de rumia (p =0.001). La relación beneficio/costo, para los tratamientos SM y SM+C, fue de 4.2 y 4.3, respectivamente. Conclusión: Bajo las condiciones de estudio, las dos estrategias de suplementación evaluadas mejoraron el desempeño de los animales y presentaron viabilidad económica, en especial la oferta simultanea de mezcla mineral y de alimento balanceado.

Introduction: In Colombia, buffalo production has been growing significantly. Buffaloes adapt well to alluvial areas, which are characterized by the presence of native grass species, such as Hymenachne amplexicaulis. Objective: To evaluate the effect of different strategies of supplementation on the productive performance and ingestive behavior of calves managed in pastures with a predominance of Hymenachne amplexicaulis. Materials and methods: Thirty Murrah calves, weighing 67.5 (SD=2) kg and 4 months old, were randomly assigned to three treatments: SS: no supplementation; SM: mineral supplement and SM+C: mineral supplement + concentrate (0.5% body weight). The animals grazed in meadows with a predominance of Hymenachne amplexicaulis, and at night they received the supplement in pens. The variables studied were weight gain and time spent on different eating behaviors, in addition to biting rate and feeding stations. Results: The mineral mixture consumption was 33.8 g in the SM and SM+C groups, and the balanced consumption was 149.3 g. The supply of SM and SM+C produced an extra increase in weight gain of 68 and 227 g, respectively (p < 0.001). Supplemented animals grazed longer (p<0.001) and ruminated less time (p =0.001). The benefit/cost ratio for the SM and SM+C treatments was 4.2 and 4.3, respectively. Conclusion: Under the study conditions, the supplementation strategies evaluated improved the performance of the animals and presented economic viability, especially the simultaneous offer of mineral mix and balanced feed.

Desenvolvimento de bebidas lácteas probióticas adicionadas de polpa de morango e suplementadas com L-triptofano elaboradas a partir de leite bubalino e bovino / Development of probiotic dairy beverages with strawberry pulp and supplemented with L-tryptophan from buffalo and bovine milk

O objetivo deste estudo foi desenvolver uma formulação de bebida láctea bubalina probiótica adicionada de polpa de morango, comparando os efeitos do uso do leite de búfala e de vaca na elaboração dos produtos e verificando a possibilidade de suplementação com triptofano nos produtos lácteos probióticos. Como primeira etapa do trabalho, bebidas lácteas probióticas foram elaboradas a partir de leite bubalino e bovino, fermentadas com Streptococcus thermophilus TA040, Lactobacillus bulgaricus LB340 e Lactobacillus acidophilus La5, e formuladas com 0, 25 e 50% de soro em sua formulação. As bebidas foram avaliadas quanto à cinética de fermentação das culturas láticas utilizadas, ao teor de proteína, gordura e sólidos totais não gordurosos, pós-acidificação, viabilidade das culturas fermentadoras e sua capacidade de sobrevivência ao estresse gastrointestinal in vitro. As bebidas lácteas bubalinas apresentaram resultados superiores as bebidas bovinas. O uso do leite de búfala na elaboração das bebidas lácteas promoveu benefícios quanto as culturas láticas presentes nos produtos, exercendo efeito protetivo e influindo na preservação da viabilidade das bactérias ao longo do armazenamento refrigerado e durante a simulação do estresse gastrointestinal in vitro. As bebidas lácteas elaboradas com 25% apresentaram os resultados mais próximos aos obtidos pelos produtos controle, sem adição de soro, sendo selecionadas para a segunda parte do estudo. Nesta etapa, as formulações de bebida láctea com 25% de soro, foram acrescidas de um preparado com polpa de morango e bebidas sem adição da fruta, utilizadas como controle. As bebidas lácteas bubalinas frutadas, apresentaram menor teor de gordura e melhores características reológicas, com maior viscosidade e consistência do que os produtos controle, sem afetar a pós-acidificação, o perfil de ácido graxo, assim como, a viabilidade e a resistência às condições de estresse gastrointestinal in vitro das culturas fermentadoras. A avaliação da possibilidade de suplementar lácteos probióticos com triptofano foi realizada em conjunto com a Universidade de Milão. Para isso, iogurtes probióticos receberam adição de triptofano antes ou após a fermentação, sendo avaliados com relação ao perfil de pós-acidificação, quantidade de triptofano nos produtos, número de células viáveis por plaqueamento e citometria de fluxo ao longo do armazenamento a 25° e 4°C. Complementarmente, a influência da presença do triptofano no crescimento e produção de compostos antimicrobianos pelas culturas láticas, também foi avaliada. A adição de triptofano após a fermentação dos iogurtes, que foram armazenados sob refrigeração (4°C), além de não afetar a pós-acidificação dos produtos, apresentou benefícios quanto a viabilidade L. acidophilus, redução do dano e aumento do número de células vivas, promovendo teor maior do aminoácido nos iogurtes. A presença do triptofano nos meios de cultivo, também influenciou de forma positiva o crescimento de S. thermophilus e L. acidophilus, melhorando o desenvolvimento das bactérias durante a fermentação e influindo em uma maior atividade antilistérica por parte do S. thermophilus. Diante da influência positiva da aplicação do leite de búfala na elaboração das bebidas lácteas, assim como, a adição do triptofano em iogurtes probióticos, a suplementação do aminoácido em bebidas lácteas bubalinas frutadas permitiria a obtenção de um produto funcional, onde seus benefícios estariam relacionados tanto ao consumo do probiótico presente no produto quanto a complementação de triptofano na dieta do consumidor

The aim of this study was to develop a formulation of probiotic buffalo dairy beverage added with strawberry pulp, comparing the effects of using buffalo and cow's milk in the preparation of products and verifying the possibility of tryptophan supplementation in probiotic dairy products. As a first stage of the work, probiotic dairy beverages were made from buffalo and bovine milk, fermented with Streptococcus thermophiles TA040, Lactobacillus bulgaricus LB340 and Lactobacillus acidophilus La5, and formulated with 0, 25 and 50% whey in their formulation. The beverages were evaluated for the fermentation kinetics of the used lactic cultures, the levels of protein, fat and total no fat solids, post-acidification, fermenting cultures viability and their ability to survive gastrointestinal stress in vitro. Buffalo milk use in dairy beverages production promoted benefits regarding the lactic cultures present in the products, exerting a protective effect and influencing the viability preservation of bacteria during the cold storage and simulation of gastrointestinal stress in vitro. Dairy beverages made with 25% whey addition showed results similar to those obtained by the control products, without whey addition, being selected for the second part of the study. In this part, the dairy beverages formulations with 25% whey, were added with a preparation were added with a strawberry pulp preparation and dairy beverages without added fruit, used as a control. Fruity bubaline dairy beverages had lower fat content and better rheological characteristics, with higher viscosity and consistency than control products, without affecting post-acidification, fatty acid profile, as well as viability and resistance to in vitro gastrointestinal condition of fermented cultures. The possibility of supplementing probiotic dairy products with tryptophan was evaluated in partnership with the University of Milan. For this, probiotic yogurts received the addition of tryptophan before or after fermentation, being evaluated in relation to the post-acidification profile, tryptophan amount in the products, viable cell number per plating and flow cytometry during storage at 25°C and 4°C. In addition, the influence of the tryptophan presence on the growth and production of antimicrobial compounds by lactic cultures was also evaluated. The addition of tryptophan after the yogurt fermentation, which were stored under refrigeration (4°C), in addition to not affecting the post-acidification of the products, showed benefits to the viability of L. acidophilus, reduced the damage and increased the number of cells promoting higher amino acid content in yogurts. Tryptophan presence in the culture media also positively influenced the growth of S. thermophiles and L. acidophilus, improving the development of bacteria during fermentation and influencing better antilisteric activity in the part of S. thermophiles. In view of the buffalo milk positive influence observed after the application in dairy beverage preparation, as well as the addition of tryptophan in probiotic yoghurts, amino acid supplementation in fruity buffalo dairy beverages would allow to obtain a functional product, where its benefits would be related both to the consumption of the probiotic present in the product as to the supplementation of tryptophan in the consumer's diet

Deciphering the functional role of EGR1 in Prostaglandin F2 alpha induced luteal regression applying CRISPR in corpus luteum of buffalo

BACKGROUND: PGF2α is essential for the induction of the corpus luteum regression which in turn reduces progesterone production. Early growth response (EGR) proteins are Cys2-His2-type zinc-finger transcription factor that are strongly linked to cellular proliferation, survival and apoptosis. Rapid elevation of EGR1 was observed after luteolytic dose of PGF2α. EGR1 is involved in the transactivation of many genes, including TGFß1, which plays an important role during luteal regression. METHODS: The current study was conducted in buffalo luteal cells with the aim to better understand the role of EGR1 in transactivation of TGFß1 during PGF2α induced luteal regression. Luteal cells from mid stage corpus luteum of buffalo were cultured and treated with different doses of PGF2α for different time durations. Relative expression of mRNAs encoding for enzymes within the progesterone biosynthetic pathway (3ßHSD, CYP11A1 and StAR); Caspase 3; AKT were analyzed to confirm the occurrence of luteolytic event. To determine if EGR1 is involved in the PGF2α induced luteal regression via induction of TGFß1 expression, we knocked out the EGR1 gene by using CRISPR/Cas9. RESULT: The present experiment determined whether EGR1 protein expression in luteal cells was responsive to PGF2α treatment. Quantification of EGR1 and TGFß1 mRNA showed significant up regulation in luteal cells of buffalo at 12 h post PGF2α induction. In order to validate the role of PGF2α on stimulating the expression of TGFß1 by an EGR1 dependent mechanism we knocked out EGR1. The EGR1 ablated luteal cells were stimulated with PGF2α and it was observed that EGR1 KO did not modulate the PGF2α induced expression of TGFß1. In PGF2α treated EGR1 KO luteal cell, the mRNA expression of Caspase 3 was significantly increased compared to PGF2α treated wild type luteal cells maintained for 12 h. We also studied the influence of EGR1 on steroidogenesis. The EGR1 KO luteal cells with PGF2α treatment showed no substantial difference either in the progesterone concentration or in StAR mRNA expression with PGF2α-treated wild type luteal cells. CONCLUSION: These results suggest that EGR1 signaling is not the only factor which plays a role in the regulation of PGF2α induced TGFß1 signaling for luteolysis.

Gastrointestinal parasitic infections of buffaloes (Bubalus bubalis) in Sarawak Borneo: Prevalence, risk factors, and farming practices

@#The objectives of this study were to investigate the prevalence and associated risk factors for gastrointestinal (GI) parasites in buffaloes from various areas of Sarawak, and to assess current management practices of GI parasites among farmers. Faecal samples were collected from 15 farms and 129 animals, as well as data on farm and animal-based characteristics. A total of 129 faecal samples were examined for GI parasites using a modified McMaster and sedimentation. Association between potential risk factors and the prevalence of GI parasites was investigated using Chi-square statistic. The prevalence of Paramphistomum sp., strongyles, and coccidia were 75.2% (95% CI±7.5), 52.7% (95% CI±8.6) and 48.1% (95% CI±8.6), respectively. Farms which had a grazing area less than 50 acres in size had significantly higher prevalence of strongyles (70.5%, χ² = 8.34, P = 0.004) and paramphistomes (88.6%, χ² = 6.46, P = 0.01) relative to farms with a larger grazing area (43.5% and 68.2%, respectively). Prevalence of strongyles was lower in farms that did not implement a cut- and-carry system (45.6%, χ² = 4.17, P = 0.04) in comparison to those that did (64%). The prevalence of paramphistomes was higher on farms with more than 40 animals (80.6%, χ² = 3.18, P = 0.05) relative to farms with fewer animals. The majority of farmers surveyed (67.9%) showed awareness of GI parasite infection and reported that they recognized the associated symptoms. Most farmers practised deworming, and ivermectin was the most commonly used anthelminthic (60.4%); only 1.9% of farmers used albendazole. Overall this study revealed a high prevalence of GI parasites in buffalo in Sarawak. Although farmers report they are aware of parasitic diseases, further education is still required. This could include how they can successfully implement on-farm changes to reduce the prevalence of GI parasites in their herds.

Effect of Milk Fermented with Lactobacillus acidophilus NCDC15 on Nutrient Digestibility, Faecal Biomarkers and Immune Response in Murrah calves

Abstract In neonates, rapid change in diet imbalances gut health allowing colonization of opportunistic pathogens that confer harmful effects on animal health causing reduced digestion and malabsorption of nutrients. In this milieu, probiotic feeding can be a promising approach in promoting animal health and stabilization of gastrointestinal microbiota. Hence, the present study was designed to investigate the effect of Lactobacillus acidophilus NCDC15 enriched fermented milk on nutrient digestibility, faecal biomarkers and immune response in Murrah buffalo calves. Twenty-four, neonatal calves (5-7 days) were randomly allocated into four groups for 90 days. The control group (CT) was provided a basal diet of calf starter and green fodder (maize and jowar), without any probiotic fermented milk (PFM) supplementation. Basal diet was supplemented with probiotic fermented milk at 100, 200 and 300 mL/calf/day, in PFM100, PFM200 and PFM300 groups, respectively. Nutrient digestibility remained similar among the groups. Faecal acetate was higher (P<0.05) in PFM200 and PFM300, while, faecal butyrate was increased (P<0.05) with all levels of probiotic supplementation than control. Faecal Lactobacillus and Bifidobacterium count were increased (P<0.05) with a concomitant reduction in coliform population (P<0.05) among all the treatments. Cell-mediated and humoral immune response were higher (P<0.001) in PFM200 and PFM300 than CT. Overall, it can be concluded that inclusion of Lactobacillus acidophilus NCDC15 in the form of fermented milk upto 300 mL/calf/day improved immunity and faecal biomarkers in Murrah buffalo calves without any adverse effect on nutrient utilization which may positively impact growth performance in Murrah buffalo calves.

Sensory acceptance and physico-chemical composition of mixed minas frescal cheese / Aceitação sensorial e composição físico-química de queijo minas frescal misto

Buffalo milk presents higher proteins and lipids concentration than cow milk, which provides a better yield in cheese-making production and products with considerable sensory approval. However, buffalo milk is not regularly available throughout the year due to different handling conditions. Thus, to guarantee the supply of buffalo milk dairy products during the year, the addition of bovine milk is an alternative. Therefore, this study aimed to test the effect of a buffalo and bovine milk mixture for the Minas Frescal cheese elaboration by physico-chemical, sensory analysis and obtaining yield. The raw material was analysed for the parameters of acidity, density, fat, total solids and solid-not-fat. There were 3 Minas Frescal cheeses elaborated from 3 formulations: 100% buffalo milk, 100% bovine milk and the mixture of 50% of each milk. The cheeses were submitted to the Gravimetric and Gerber methods to obtain values of moisture and fat, as well as to obtain, indirectly, fat in dry matter. For bovine and buffalo milk, the parameters evaluated (acidity, density, fat, total solids and solids-not-fat) complied with Brazilian legislation and parameters described in the literature. For the bovine, mixed and buffalo fresh cheeses, values were obtained, respectively, for moisture (74.04, 60.93 and 63.61), fat in dry matter (44.35, 42.23 and 46.03) and cheese yield (27, 20.8 and 24.2), indicating a higher yield for the bovine Minas Frescal cheese and higher fat content for the buffalo cheese. The overall acceptance of the mixed Minas Frescal cheese was significantly superior to the bovine and buffalo cheese. The parameters of colour, appearance, texture, flavour and overall acceptance were above 8 points in the hedonic scale ('moderately liked' to 'extremely liked'), and the aroma attribute scored was above 7 ('I enjoyed regularly' to 'moderately liked'). Therefore, the elaboration of Minas Frescal cheese from the mixture of 2 matrices (buffalo and bovine milk) demonstrated technological viability with the potential to meet the demands of the consumer market.

O leite de búfala possui maior porcentagem de todos os componentes tornando-o uma matéria-prima adequada com potencial para fornecer um maior rendimento e qualidade nutricional na produção de queijo. No entanto, o leite de búfalo não está disponível regularmente ao longo do ano devido a diferentes condições de manejo. Assim, para garantir a oferta de produtos lácteos com leite de búfala independentemente da época, a adição de leite bovino pode ser vista como uma alternativa. Logo, este trabalho teve como objetivo testar o efeito da mistura do leite de búfalo e bovino na elaboração do queijo Minas Frescal por meio de análises físico-químicas e sensoriais e obtenção do rendimento economico. A matéria-prima foi analisada quanto aos parâmetros: acidez titulável, densidade, gordura, sólidos totais e sólidos totais desengordurados. Três variedades de queijo Minas Frescal foram elaboradas a partir de três formulações: 100% de leite de búfala, 100% de leite bovino e da mistura de 50% de cada leite. Os queijos foram submetidos aos métodos Gavimetric e Gerber para obtenção de umidade e gordura, bem como, indiretamente, para obtenção de matéria gorda no extrato seco. Para o leite bovino e de búfala, os parâmetros avaliados (acidez titulável, densidade, gordura, sólidos totais e sólidos totais desengordurados) estavam de acordo com a legislação brasileira e parâmetros descritos na literatura. Para os queijos bovino, misto e de búfala, foram obtidos, respectivamente, para: umidade (74,04; 60,93 e 63,61), gordura na matéria seca (44,35; 42,23 e 46,03) e rendimento econômico (27, 20,8 e 24,2) indicando maior rendimento o queijo Minas Frescal bovino e maior teor de gordura para o queijo búfalo. A aceitação global do queijo Minas Frescal misto foi significativamente superior ao do queijo bovino e bubalino. Os parâmetros de cor, aparência, textura, sabor e aceitabilidade global foram acima de 8 pontos (entre "gostei moderadamente" e "extremamente gostei") e o atributo aroma obteve pontuação acima de 7 ("gostei regularmente" para "gostei moderadamente"). Portanto, a elaboração do queijo Minas Frescal da mistura de leite bovino ao búfalo demonstrou potencial para atender as demandas do mercado consumidor.

Comparative Evaluation of the Efficacy of Herbal Regimen for the Treatment of Post-partum Anestrus in Water Buffaloes

A study was undertaken to compare the efficacy of combinations of different brands of herbal uterine cleansers, estrus inducers and mineral supplements in the treatment of anestrus in water buffaloes. Thirty she-buffaloes with a history of anestrus were randomly assigned to either of five equal groups. Group T0 served as untreated control, Group T1 was treated with a combination of Exapar bolus (M/s Ayurvet Limited, India), Janova capsule (M/s Ayurvet Limited, India) and Mintrus caplet (M/s Ayurvet Limited, India). Group T2 was treated with AV/UTB/26 bolus (M/s Ayurvet Limited, India), AV/OIP/22 powder (M/s Ayurvet Limited, India) and Mintrus caplet. Group T3 was treated with a combination of Brands A, B and C. Group T4 was treated with a combination of Brands X, Y and Z. The best estrus response was recorded in the Group T2 and the highest conception rates in Groups T2 and T4. The time interval between treatment and onset of estrus was significantly lowest (P<0.05) in Group T1 as compared to the other groups

Caracterización de decomisos de vísceras rojas en un frigorífico de exportación en Santander - Colombia / Characterising red viscera confiscation in an export slaughterhouse in Santander, Colombia / Caracterização de confiscos de vísceras vermelhas num frigorífico de exportação em Santander - Colômbia

Resumen A nivel mundial el decomiso de vísceras rojas es una de las principales causas de pérdidas económicas en la comercialización de productos cárnicos. El objetivo del presente estudio fue determinar las principales causas y variables epidemiológicas asociadas al decomiso de vísceras rojas en el FRIGORÍFICO RÍO FRÍO (FRF), del municipio de Floridablanca, Santander y estimar el valor de las pérdidas económicas asociadas a esta actividad. Entre 9 de noviembre al 9 de diciembre de 2018, se realizó un estudio transversal en el total de animales sacrificados en el FRF, a los cuales se les recolectó información epidemiológica posterior a la inspección sanitaria realizada por funcionarios del INVIMA. Las principales causas y variables epidemiológicas asociadas al decomiso de vísceras rojas se estimaron a partir de análisis de frecuencias, mientras el valor de las pérdidas económicas, a través de un análisis económico en el mercado nacional. Se sacrificaron un total de 8671 bovinos, y 226 búfalos, en los cuales se realizó el decomiso de vísceras rojas en el 15,8% y 26,5% de los animales, respectivamente. En los bovinos los principales órganos decomisados fueron pulmón, hígado y bazo con porcentajes del 87,4%, 11,1% y 0,6%, respectivamente. Mientras que, en los búfalos, fueron el pulmón, hígado y riñón con porcentajes del 88,3%, 8,3% y 1,7%, respectivamente. Las principales causas de decomiso de pulmón en ambas especies fueron la broncoaspiración (86,6%) y enfisema (9,4%), mientras para el hígado fueron los abscesos (40,5%) y la distomatosis hepática (28,8%) en los bovinos, hígado graso (80,0%) y cirrosis (20,0%) en los búfalos. Para el caso de los bovinos, hubo diferencias significativas en la proporción de animales con distomatosis hepática entre los diferentes municipios de Santander (χ² = 50.03, P < 0.05). Las mayores pérdidas económicas durante el periodo de estudio se deben al decomiso de pulmón e hígado con valores de 4886 y 2639 USD. En conclusión, el pulmón fue el órgano decomisado con mayor frecuencia en ambas especies, principalmente asociado a la broncoaspiración. Futuras estrategias de intervención, con el fin de reducir la broncoaspiración en los animales que ingresan a la planta de beneficio, e interrumpir el ciclo de vida de Fasciola hepatica en algunos municipios de Santander, son necesarios para disminuir el nivel de pérdidas económicas a lo largo de la cadena productiva.

Abstract Red viscera confiscation represents a significant cause of economic loss for the meat products' trade. This study was aimed at determining the causes and epidemiological variables associated with red viscera confiscation in a slaughterhouse (FRIGORÍFICO RÍO FRÍO (FRF)) located in Floridablanca, in Colombia's Santander department, and estimating the value of the economic losses associated with such confiscation. A cross-sectional study was carried out between November and December 2018 based on epidemiological information regarding all animals slaughtered at the FRF recorded after routine health inspection by Colombian Food and Drug Surveillance Institute (INVIMA) officials. Frequency analysis was used for estimating the main causes of and the epidemiological variables associated with red viscera confiscation whilst economic losses were estimated by national market analysis. It was found that red viscera were confiscated in 15.8% of 8,671 sacrificed cattle compared to 26.5% for 226 buffaloes. The lungs (87.4%), liver (11.1%) and spleen (0.6%) were the main organs seized for cattle and lungs (88.3%), liver (8.3%) and kidneys (1.7%) for buffaloes. Bronchoaspiration (86.6%) and emphysema (9.4%) were the main causes of confiscation regarding the lungs in both species while for the liver these were abscesses (40.5%) and hepatic dystomatosis (28.8%) in cattle and fatty liver (80.0%) and cirrhosis (20.0%) in buffaloes. There were significant differences regarding the percentage of cattle having hepatic dystomatosis amongst municipalities in the Santander department (χ²=50.03, p<0.05). Lung (4,886 USD) and liver confiscation (2,639 USD) led to the greatest economic losses; the lungs were the main organ confiscated in both species, mainly being associated with bronchoaspiration. Future intervention strategies are needed to reduce the livestock industry's financial losses, aimed at reducing bronchoaspiration in animals in the slaughterhouse and interrupting Fasciola hepatica's life-cycle in some municipalities in the Santander department.

Resumo Globalmente, os confiscos de órgãos e vísceras vermelhas é uma das principais causas de perdas econômicas na comercialização de produtos à base de carne. O objetivo deste estudo foi determinar as principais causas e variáveis epidemiológicas associadas aos confiscos de vísceras vermelhas num frigorífico (FRIGORÍFICO RÍO FRÍO (FRF)), do município de Floridablanca, Santander, e estimar o valor das perdas econômicas associadas a esta atividade. Entre 9 de novembro e 9 de dezembro do 2018, foi realizado um estudo transversal sobre o total de animais abatidos no FRF, nos quais a informação epidemiológica era diligenciada depois da inspeção sanitária realizada pelos funcionários do INVIMA. As principais causas e variáveis epidemiológicas associadas ao confisco de vísceras vermelhas foram estimadas a partir da análise de freqüência, e o valor de perdas econômicas, através de uma análise no mercado nacional. Foram abatidos 8.671 bovinos e 226 búfalos, nos quais 15,8% e 26,5% tiveram confiscos de vísceras vermelhas, respectivamente. Em bovinos, as principais vísceras confiscadas, foram pulmões, fígado e baço com percentagens de 87,4%, 11,1% e 0,6%, respectivamente. Nos búfalos, foram pulmões, fígado e rim com porcentagens de 88,3%, 8,3% e 1,7%, respectivamente. Para as duas espécies, broncoaspiração (86,6%) e enfisema (9,4%) foram as principais causas de confiscos dos pulmões. Para o fígado, foram os abcessos (40,5%) e fasciolose (28,8%) em bois, esteatose hepática (80,0%) e cirrose (20,0%) em búfalos. No caso dos bois, houve diferenças significativas na proporção de animais com fasciolose nos diferentes municípios do Santander (χ² = 50,03, P <0,05). As maiores perdas econômicas durante o período de estudo correspondem ao confisco de pulmão e fígado com valores de 4886 e 2639 USD. Em conclusão, o pulmão foi a principal víscera confiscada em ambas espécies, principalmente associados a broncoaspiração. Estratégias futuras de intervenção, com o objetivo de reduzir a broncoaspiração, e interromper o ciclo de vida da Fasciola hepatica em alguns municípios do Santander são necessários para reduzir as perdas econômicas na cadeia de produção.

Potential of microbial inoculum from buffalo feces in activating lignite coal bed methane

Aim: This research aimed to study the ability of microbial consortium from buffalo feces to produce coal bed methane (CBM) in lignite coal through in-vitro technique.Methodology: This study was carried out in 2 stages: In first stage, microbes activated using in-vitro technique, and at second stage volatile fatty acids (VFA) and coal bed methane production were estimated. The nutrition source for inoculum activation was of three types: 100% concentrate: 70% concentrate: 30% grass and 30% concentrate: 70% grass. Biogas digester in laboratory-scale utilized 100 ml serum bottle filled with 70 ml, 98-5 media and 7 g coal. Microbial inoculum was inoculated on digester using a 10 ml syringe and incubated at 39oC. Parameters observed to measure the quality of inoculum were total number of anaerobic microbes, kinetics, and biogas production during fermentation. Complete Randomized Design with two factorial consisted of incubation period as factor A and inoculum dosages as factor B. Results: The activation process of inoculum from buffalo feces was required to produce coal bed methane in anaerobic digestion. During activation process, the microbes from buffalo feces in a mixture of 70% concentrate and 30% grass at 48 hr observation could produce total gas approximately 22.5 ml at 48 hr observation. Addition of activated buffalo feces anaerobic inoculum on the anaerobic digestion as much as 6% produced the highest number of anaerobic bacteria, and VFA accounted for 31 x 1010 CFU ml-1 and 171.7 mM, respectively. Meanwhile, the highest methane production reached 128.61 ml on adding 6% inoculum

Isolation of Pseudomonas aeruginosa from Bovine Mastitic Milk Sample Along with Antibiogram Study

A total of 4378 cattle and buffalo were screened during period of study (July 2018-June 19). Out of which 27 Pseudomonas aeruginosa were isolated. The isolates were confirmed phenotypically based on pigment production on nutrient agar. These were then confirmed by PCR amplification of species specific oligonucleotide sequences. All the 27 isolates amplified 956bp amplicon 16srRNA Pseudomonas aeruginosa species specific nucleotide sequence. The isolates were also checked for exo and aglD virulence associated genes. All of them amplified 540bp and 313bp amplicon of exo gene and aglD gene. All the isolates were subjected to antibiotic sensitivity testing. Most of the isolates showed highest sensitivity for levofloxacin, streptomycin and enrofloxacin followed by gentamicin, moxifloxacin and amikacin. Neomycin, cefoperazone and ceftriaxone were intermediate in action

Comparative Efficacy of PercollTM Discontinuous Density Gradient Centrifugation and Glass Wool Filtration Techniques for Spermatozoa Selection in Buffalo (Bubalus bubalis)

Dead and damaged spermatozoa cells present universally in the ejaculates of all eutherian mammals exert toxic effect on contemporary healthy cells mostly through generation of excessive free radicals. This is much more evident during extended period of processing, resulting in poor ejaculate quality. The solution lies in depletion of dead/damaged spermatozoa from the neat ejaculates itself. Thus the objective of the study was to evaluate the efficiency of the protocols such as discontinuous PercollTM density gradient centrifugation (PDGC) and glass wool filtration (GWF) for depletion of dead/damaged spermatozoa from fresh semen in buffalo. Random ejaculates (n=6) of Murrah buffalo bulls were divided into two aliquots after quality assessment: PDGC and GWF protocols (Group I and II, respectively). At the end of the purification protocol, efficiency of the protocols in depleting dead/damaged spermatozoa as reflected by certain quality parameters were evaluated. The mean efficiency (%) of purification protocols based on recovery of spermatozoa was 44.68 and 40.02% for PDGC and GWF, respectively. Moreover significantly (p<0.05) greater values for quality parameters was observed in the Group II (26.4+6.8 vs 68.8+4.4 for acrosome integrity (%); 12.68+6.6 vs 57.7+7.5 for functional plasma membrane integrity (%); 20.3+5.8 vs 80.75+6.7 for viability (%) in Group I and II, respectively). It was concluded that GWF is a better technique than PGDC to filter out dead/damaged spermatozoa from fresh semen with improvement in semen quality and can be a valuable tool in assisted reproductive technology

Cotylophoron marajoensis n. sp. (Digenea: Paramphistomidae) a parasite of Bubalus bubalis on Marajó Island, Pará, Brazilian Amazon / Cotylophoron marajoensis n. sp. (Digenea: Paramphistomidae) parasito de Bubalus bubalis na Ilha de Marajó, Pará, Amazônia Brasileira

Abstract The genus Cotylophoron belongs to the Paramphistomidae family and its definitive hosts are ruminants in general. This work describes the presence of a new species of the gender, a parasite in the rumen and reticulum of Bubalus bubalis, on Marajó Island in the Eastern Brazilian Amazon, using of light microscopy, scanning electronic microscopy and molecular biology techniques. One hundred and ten animals were analyzed, of which 4.54% were parasitized by flukes in their adult forms. The helminths were found fixed to the ruminal mucosa and present Liorchis-type pharynx, Cotylophoron-type genital sucker, oblique testicles larger than the ovary, uterus in rings full of eggs and Cotylophoron-type acetabulum. These morphologic characters do not fit into any previously described species. Thus, it is proposed that this is a new species in the genus Cotylophoron. The present work expands the record of parasitism by helminths in Bubalus bubalis, this being the first record of trematoda from the genus Cotylophoron for this host in the Brazilian Amazon.

Resumo O gênero Cotylophoron pertence à família Paramphistomidae e possui como hospedeiros definitivos ruminantes em geral. Este trabalho descreve a presença de uma espécie nova do gênero, parasito do rúmen e retículo de Bubalus bubalis, na Ilha de Marajó, Amazônia oriental brasileira, a partir das técnicas de microscopia de luz, microscopia eletrônica de varredura e biologia molecular. Foram analisados 110 animais, dos quais 4,54% estavam parasitados por trematódeos na sua forma adulta. Os helmintos foram encontrados fixados à mucosa ruminal, apresentando faringe do tipo Liorchis, ventosa genital do tipo Cotylophoron, testículos oblíquos maiores que o ovário, útero em alças repleto de ovos, e acetábulo do tipo Cotylophoron. Estes caracteres morfológicos não se enquadram em nenhuma espécie previamente descrita. Assim, propõe-se uma nova espécie ao gênero Cotylophoron. O presente trabalho amplia o registro do parasitismo por helmintos em Bubalus bubalis, sendo este o primeiro registro de trematódeos do gênero Cotylophoron nesse hospedeiro para a Amazônia brasileira.

Effects of different culture systems on the culture of prepuberal buffalo (Bubalus bubalis) spermatogonial stem cell-like cells in vitro