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.

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

The objective of the current study was to investigate the synergistic impact of α-Tocopherol and α-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 + 50µM α-Tocopherol, GROUP 3: MM + 100 µM ALA + 100µM α-Tocopherol, GROUP 4: MM + 100 µM ALA + 200 µM α-Tocopherol and GROUP 5: MM + 100 µM ALA + 300 µM α-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 Tyrode's Albumin Lactate Pyruvate (TALP) medium for about 20 h and cultured in synthetic oviductal fluid (SOF) medium to determine effect of α-Linolenic acid (100 µM) and α-Tocopherol in IVM medium on IVC of presumptive zygotes. To study the effect of α-Linolenic acid (100 µM) in IVM media and increasing concentration of α-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 α-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 α-tocopherol in maturation media having α-Linolenic acid and/or in embryo culture media did not further enhance in vitro maturation of oocyte or embryo production.

Demographics of Cystic Echinococcosis Patients Treated Surgically in Lahore, Pakistan: A Single Centre Study from 2007 - 2018.

Cystic echinococcosis (CE) is a zoonotic disease caused by the larval stage of Echinococcus granulosus tapeworms. These parasites have a worldwide geographic distribution and pose a serious threat to livestock industry as well as human health in the endemic areas. CE is widely distributed in Pakistan. However, very few reports are available related to the regional transmission of E. granulosus. A retrospective analysis was conducted of surgically confirmed CE patients who were treated at Shoukat Khanum Memorial Cancer Hospital and Research Centre in Lahore, Punjab Province, Pakistan from 2007 - 2018. In total, 536 CE patients were evaluated during the study period. Cases originated from the provinces of Khyber Pakhtunkhwa (n=336), Punjab (n=147), Baluchistan (n=18), Sindh (n=3), Islamabad (n=2), Gilgit Baltistan (n=1), and Azad Jammu and Kashmir (n=1). An additional 28 cases were from Afghanistan. The highest number of CE cases was reported in 2013 (n=90). Females made up a larger proportion of cases (n=310; 57.8 %) than males (n=226; 42.2 %). Most patients were members of the Pashtun (n=197; 36.7 %), Hindku (n=142; 26.5 %), and Punjabi (n=118; 22.0 %) ethnic groups. The largest number of cysts was obtained from the liver (137/536; 25.6 %). This study showed that CE is likely present throughout Pakistan. In order to control the disease, a comprehensive control program and regional surveillance are needed.

Poverty-associated emerging infection of Cystic Echinococcosis in population of Northern Pakistan: A hospital based study.

Cystic Echinococcosis (CE) is one of the most important zoonotic parasitic diseases in human, livestock, and wildlife globally. The prevalence of CE depends upon human behavioral risk factors, the diversity and ecology of animal host interactions and the genetic diversity within Echinococcus species which differ in their zoonotic potential and pathogenicity. It is a neglected, economic and socio-cultural problem in Pakistan. The available data about the incidence of CE is very limited and no extensive study has been reported in Pakistan. The current study was aimed to analyze the hospital reported cases of CE and the associated risk factors related to the incidence of CE. The hospital-based data of CE for the time period of January 2012-December 2017 was collected from Islamabad, Rawalpindi and Peshawar. The data covered demographic characteristics including age, gender, and cyst localization of infected individuals and socioeconomic determinants. The data was analyzed based upon different risk factors along with the different socioeconomic parameters that has an important impact on the distribution of disease. A total of 228 cases were presented in the selected hospitals of different cities during the study period. Out of total 228 patients, 59.21% were males and 40.78% were females (P<0.001). Most infections have been recorded in young adults (>20-30) showing 22.8% of total infected individuals followed by children (0-10) showing 10.5% (n=24), respectively (P<0.001). Liver was the most vulnerable organ (58.77%, n=134) followed by lungs (14.47%, n=33) (P<0.001). The infection was higher among rural communities (84.2%) than urban (12.8%) (P<0.001). Socioeconomic and demographic factors had an important impact on the intensity of disease (P<0.001). The occurrence of cases in children and young adults was an important finding as it indicated an active transmission of the parasite in Pakistan along with the poverty index. Emergence of echinococcosis in Pakistan showed that emerging health issues in Pakistan could bring the disease to limelight for future research. This finding, together with the fact that 1 hospital reported 214 cases over 6 years 325 underlines the need for a program for prevention/control of this disease in Pakistan. The timely measure needs to be taken to hamper the disease development and establishment. In order to control the disease, complete surveillance should be done which in turn weighs down the disease progress.

Poverty-associated emerging infection of Cystic Echinococcosis in population of Northern Pakistan: A hospital based study

@#Cystic Echinococcosis (CE) is one of the most important zoonotic parasitic diseases in human, livestock, and wildlife globally. The prevalence of CE depends upon human behavioral risk factors, the diversity and ecology of animal host interactions and the genetic diversity within Echinococcus species which differ in their zoonotic potential and pathogenicity. It is a neglected, economic and socio-cultural problem in Pakistan. The available data about the incidence of CE is very limited and no extensive study has been reported in Pakistan. The current study was aimed to analyze the hospital reported cases of CE and the associated risk factors related to the incidence of CE. The hospital-based data of CE for the time period of January 2012-December 2017 was collected from Islamabad, Rawalpindi and Peshawar. The data covered demographic characteristics including age, gender, and cyst localization of infected individuals and socioeconomic determinants. The data was analyzed based upon different risk factors along with the different socioeconomic parameters that has an important impact on the distribution of disease. A total of 228 cases were presented in the selected hospitals of different cities during the study period. Out of total 228 patients, 59.21% were males and 40.78% were females (P<0.001). Most infections have been recorded in young adults (>20-30) showing 22.8% of total infected individuals followed by children (0-10) showing 10.5% (n=24), respectively (P<0.001). Liver was the most vulnerable organ (58.77%, n=134) followed by lungs (14.47%, n=33) (P<0.001). The infection was higher among rural communities (84.2%) than urban (12.8%) (P<0.001). Socioeconomic and demographic factors had an important impact on the intensity of disease (P<0.001). The occurrence of cases in children and young adults was an important finding as it indicated an active transmission of the parasite in Pakistan along with the poverty index. Emergence of echinococcosis in Pakistan showed that emerging health issues in Pakistan could bring the disease to limelight for future research. This finding, together with the fact that 1 hospital reported 214 cases over 6 years 325 underlines the need for a program for prevention/control of this disease in Pakistan. The timely measure needs to be taken to hamper the disease development and establishment. In order to control the disease, complete surveillance should be done which in turn weighs down the disease progress.

Anthelmintic properties of extracts from Artemisia plants against nematodes.

Artemisia plant genus, natural inhabitant of northern Punjab Pakistan, is well known for its anthelmintic properties; many Artemisia species have not been so far scientifically proved. The aim of this study was to assess in vitro anthelmintic activity of Artemisia indica and Artemisia roxburghiana against mixed infection of gastrointestinal nematodes in small ruminants. This study is first scientifically proven study on anthelmintic activity of A. indica and A. roxburghiana. Five different concentrations (50, 25, 12.5, 6.25 and 3.75 mg/mL) accompanied by negative control (PBS) and positive control (albendazole, 10%) were used to carry out the egg hatch inhibition assay, larval mortality assay and adult worm mortality assay. The Baermann technique was used first time in larval mortality assay and proved to be effective. The results revealed that methanolic extracts of both A. indica and A. roxburghiana, showed maximum anthelmintic activity at concentration of 50 mg/ml by egg hatch inhibition (85±21.2; 80±28.3), larvae mortality (18±2.8; 17±4.2) and adult worm mortality (8.5±2.1; 8±2.8) assays. However, at concentration of 50 mg/ml both plant extracts in comparison to albendazole showed statistically insignificant (p≤0.05) results. The A. indica showed higher anthelmintic activity at all concentrations as compared to A. roburghiana. It has been concluded both plants exhibit anthelmintic activity and further evaluation of these plants should be carried out to purify the active ingredients for anthelmintic activity. Moreover, the decoctions of these plants could be used to GINs after confirming anthelmintic properties through in vivo.

Seroprevalence of IgG and IgM antibodies and associated risk factors for toxoplasmosis in cats and dogs from sub-tropical arid parts of Pakistan.

Pet cats and dogs are an important source of human toxoplasmosis because of their intimate relationship with humans. Present study was designed to determine the prevalence and risk factors of toxoplasmosis in cats and dogs in northern sub-tropical arid region of Pakistan where no such work has been previously conducted. For this study 420 cats and 408 dogs visiting different pet clinics and veterinary hospitals were screened for the presence of anti-Toxoplasma IgG and IgM antibodies using ELISA technique. Epidemiological information regarding age, sex, area, outdoor access and hunting practice was obtained from the owners by questionnaire interview. Overall seroprevalence in cats and dogs was 26.43% (111/420) and 28.43% (116/408) respectively. IgG antibodies were found in 23.33% (98) cats and 25.49% (104) dogs while IgM antibodies were found in 3.57% (15) cats and 3.92% (16) dogs. Seroprevalence was significantly high in cats and dogs older than one year. No significant difference was recorded between males and females. Cats and dogs from rural areas showed higher prevalence. Dogs which had access to outside also showed high seroprevalence. The present study indicates that Toxoplasma gondii is widespread in pet animals in Pakistan which may have important implication for public health.

Mobilisation into cotton and spread of a recombinant cotton leaf curl disease satellite.

Analysis of a DNA beta satellite associated with a recently identified cotton leaf curl disease (CLCuD) strain indicated it to be recombinant, with most of the molecule originating from CLCuD DNA beta but with some sequence from a satellite isolated from tomato. Analysis of both archival (pre 2001) and recent cotton samples, shows the recombinant satellite is confined to a small area but was not present in cotton prior to 2001. This indicates that the recombinant DNA beta was recently mobilized into cotton, likely from tomato, and that recombination plays a role in the evolution of these satellites.

Seasonal variation in mood in African American college students in the Washington, D.C., metropolitan area.

OBJECTIVE: The authors attempted to estimate the occurrence, frequency, and pattern (winter versus summer) of seasonal affective disorder in African American college students. They hypothesized that winter seasonal affective disorder would be more prevalent than summer seasonal affective disorder. METHOD: Undergraduate and graduate college students who identified themselves as African Americans living in the Washington, D.C., metropolitan area were invited to participate in the study. The Seasonal Pattern Assessment Questionnaire was used to calculate a global seasonality score and to estimate the frequency of seasonal affective disorder and subsyndromal seasonal affective disorder. The frequency of the summer versus winter pattern of seasonality of seasonal affective disorder was compared by using multinomial probability distribution tests. The effects of gender and the awareness of seasonal affective disorder were evaluated with a two-way analysis of variance. RESULTS: Of 646 students who were invited to participate, 597 returned the questionnaires, and 537 (83.1%) fully completed them. Winter seasonal affective disorder was significantly more prevalent than summer seasonal affective disorder. The mean global seasonality score was 8.3 (SD=5.3). The majority of the subjects (80%) were not aware of the existence of seasonal affective disorder. CONCLUSIONS: The authors found that the frequency, magnitude, and pattern of seasonality of mood in African American students were similar to those previously reported in the general population at similar latitude, but that awareness of the existence of seasonal affective disorder, a condition with safe and effective treatment options, was lower.