Identification of overlay differentially expressed genes in both rats and goats with blast lung injury through comparative transcriptomics / 中华创伤杂志(英文版)

PURPOSE@#To identify the potential target genes of blast lung injury (BLI) for the diagnosis and treatment.@*METHODS@#This is an experimental study. The BLI models in rats and goats were established by conducting a fuel-air explosive power test in an unobstructed environment, which was subsequently validated through hematoxylin-eosin staining. Transcriptome sequencing was performed on lung tissues from both goats and rats. Differentially expressed genes were identified using the criteria of q ≤ 0.05 and |log2 fold change| ≥ 1. Following that, enrichment analyses were conducted for gene ontology and the Kyoto Encyclopedia of Genes and Genomes pathways. The potential target genes were further confirmed through quantitative real-time polymerase chain reaction and enzyme linked immunosorbent assay.@*RESULTS@#Observations through microscopy unveiled the presence of reddish edema fluid, erythrocytes, and instances of focal or patchy bleeding within the alveolar cavity. Transcriptome sequencing analysis identified a total of 83 differentially expressed genes in both rats and goats. Notably, 49 genes exhibited a consistent expression pattern, with 38 genes displaying up-regulation and 11 genes demonstrating down-regulation. Enrichment analysis highlighted the potential involvement of the interleukin-17 signaling pathway and vascular smooth muscle contraction pathway in the underlying mechanism of BLI. Furthermore, the experimental findings in both goats and rats demonstrated a strong association between BLI and several key genes, including anterior gradient 2, ankyrin repeat domain 65, bactericidal/permeability-increasing fold containing family A member 1, bactericidal/permeability-increasing fold containing family B member 1, and keratin 4, which exhibited up-regulation.@*CONCLUSIONS@#Anterior gradient 2, ankyrin repeat domain 65, bactericidal/permeability-increasing fold containing family A member 1, bactericidal/permeability-increasing fold containing family B member 1, and keratin 4 hold potential as target genes for the prognosis, diagnosis, and treatment of BLI.

Effects of orientation and distance of goats on blast lung injury characteristics on a plateau above 4500-meter / 中华创伤杂志(英文版)

PURPOSE@#High explosives are used to produce blast waves to study their biological effects. The lungs are considered as the critical target organ in blast-effect studies. The degree of lung hemorrhaging is related to both the explosive power and the increased lung weight. We studied the characteristics of the biological effects from an air explosion of a thermobaric bomb in a high-altitude environment and the lethality and lung injury severity of goats in different orientations and distances.@*METHODS@#Goats were placed at 2.5, 3, 4, and 5 m from the explosion center and exposed them to an air blast at an altitude of 4700-meter. A group of them standing oriented to the right side and the other group seated facing the explosion center vertically. The lung injuries were quantified according to the percentage of surface area contused, and using the pathologic severity scale of lung blast injury (PSSLBI) to score the 4 injury categories (slight, moderate, serious and severe) as 1, 2, 3, and 4, respectively. The lung coefficient (lung weight [g]/body weight [kg]) was the indicator of pulmonary edema and was related to lung injury severity. Blast overpressure data were collected using blast test devices placed at matching locations to represent loadings to goats. All statistical analyses were performed using SPSS, version 26.0, statistical software (SPSS, Inc., Chicago, IL, USA).@*RESULTS@#In total, 127 goats were involved in this study. Right-side-standing goats had a significantly higher mortality rate than those seated vertical-facing (p < 0.05). At the 2.5 m distance, the goat mortality was nearly 100%, whereas at 5 m, all the goats survived. Lung injuries of the right-side-standing goats were 1 - 2 grades more serious than those of seated goats at the same distances, the scores of PSSLBI were significantly higher than the seated vertical-facing goats (p < 0.05). The lung coefficient of the right-side-standing goats were significantly higher than those of seated vertical-facing (p < 0.05). Mortality, PSSLBI, and the lung coefficient results indicated that the right-side-standing goats experienced severer injuries than the seated vertical-facing goats, and the injuries were lessened as the distance increased. The blast overpressure was consistent with these results.@*CONCLUSION@#The main killing factors of the thermobaric bomb in the high-altitude environment were blast overpressure, blast wind propulsions and burn. The orientation and distances of the goats significantly affected the blast injury severity. These results may provide a research basis for diagnosing, treating and protecting against injuries from thermobaric explosions.

Prevalence of gastrointestinal parasite in small ruminants of District Dir Upper Khyber Pakhtunkhwa Province of Pakistan / Prevalência de parasita gastrointestinal em pequenos ruminantes do distrito Upper Dir, da província de Khyber Pakhtunkhwa, no Paquistão

Abstract The present research was planned to assess the occurrence of intestinal parasites in small ruminants of Upper Dir of Khyber Pakhtunkhwa Province of Pakistan. For this purpose, the faecal material was collected randomly with gloved fingers directly from the rectum region of sheep and goats and the faecal materials were then put in hygienic plastic bottles with 10% formalin. The overall 315 (n=184 sheep and n= 131 goats) faecal samples were collected out of 315 samples, 281 were found positive for different parasites. Patterns-wise prevalence of GI parasites of the study area was found. Overall Single parasitism 89.20% (281/315) with 94.0% (173/184) in sheep and 82.43% (108/131) in goats. Double parasitic infection in small ruminant recorded in which Fasciola+ Haemonchus. contortus in sheep were found their prevalence was 25.54% (47/184). While in goats, the double parasitic infection in which Haemonchus contortus+Trichuris spp were found and their prevalence were 23.43% (30/131). The species found in the sample of sheep were includes, i.e., Strongyloides papillosus (41.30%), Heamonchus controtus (21.73%), Trichuris ovis (17.39%), and Fasciola hepatica (13.58%), the corresponding value for goat were Strongyloides spp 33.33% (36/108), Haemonchus spp 28.70%, (27/108), Trichuris spp 25.20% (27/184) and Fasciola spp 10.68% (14/184). The sheep of the study area are more infected as compared to goats. This study suggested that gastrointestinal parasites are major health problems of small ruminants in the study area. Therefore, a comprehensive study on species of gastrointestinal parasites circulating in the area, control options, cost-effective strategies and awareness about gastrointestinal parasites among the farmers in the study area should be instituted.

Resumo A presente pesquisa foi planejada para avaliar a ocorrência de parasitas intestinais em pequenos ruminantes do distrito Upper Dir, da província de Khyber Pakhtunkhwa, no Paquistão. Para tanto, o material fecal foi coletado aleatoriamente com dedos enluvados diretamente da região do reto de ovelhas e cabras, e os materiais fecais foram colocados em garrafas plásticas higiênicas com formol a 10%. No total, 315 (n = 184 ovelhas e n = 131 cabras) amostras fecais foram coletadas; destas, 281 foram consideradas positivas para diferentes parasitas. A prevalência de padrões de parasitas GI da área de estudo foi encontrada. Parasitismo global único 89,20% (281/315) com 94,0% (173/184) em ovinos e 82,43% (108/131) em cabras. Infecção parasitária dupla em pequenos ruminantes registrada em Fasciola + Haemonchus contortus em ovinos, sua prevalência foi de 25,54% (47/184). Já em caprinos, a dupla infecção parasitária em que Haemonchus contortus + Trichuris spp foram encontrados e sua prevalência foi de 23,43% (30/131). As espécies encontradas na amostra de ovinos foram: Strongyloides papillosus (41,30%), Heamonchus controtus (21,73%), Trichuris ovis (17,39%) e Fasciola hepatica (13,58%), o valor correspondente para cabra foi Strongyloides spp. 33,33% (36/108), Haemonchus spp. 28,70%, (27/108), Trichuris sp 25,20% (27/184) e Fasciola spp. 10,68% (14/184). As ovelhas da área de estudo estão mais infectadas do que as cabras. Este estudo sugeriu que os parasitas gastrointestinais são os principais problemas de saúde dos pequenos ruminantes na área de estudo. Portanto, um estudo abrangente sobre as espécies de parasitas gastrointestinais que circulam na área, opções de controle, estratégias de baixo custo e conscientização sobre parasitas gastrointestinais entre os agricultores na área de estudo deve ser instituído.

The inhibitory effect of Cannabis Sativa L. and Morus nigra L. against lipid peroxidation in goat liver and brain homogenates / O efeito inibitório de Cannabis Sativa L. e Morus nigra L. contra a peroxidação lipídica em homogenatos de fígado e cérebro de cabra

Abstract The present study was aimed to evaluate the antioxidant potential and inhibitory effect ofCannabis sativa and Morus nigra against lipid peroxidation in goat brain and liver homogenates. The formation of free radicals, highly reactive oxygen species (ROS) and reactive nitrogen species (RNS) is a normal metabolic process for cellular signaling and countering the antigens. However, they may cause serious damage if they produced at amplified tolls. In addition, metabolic disorders also serve as sources of these reactive species. Although the issue can be addressed through supplements and other phytochemicals. In this study, two plant species were evaluated for their biological potential by employing a spectrum of antioxidant assays. The antioxidant activity was performed by lipid peroxidation assay. The water extract prepared from leaves of Cannabis sativa and Morus nigra showed significant (P<0.05) inhibition as compared to control i.e., 522.6±0.06 and 659.97±0.03 µg/mL against iron-induced lipid peroxidation in goat brain homogenate while the inhibitions were 273.54±0.04 and 309.18±0.05 µg/mL against nitroprusside induced lipid peroxidation of the brain. The iron and nitroprusside induced lipid peroxidation was also significantly inhibited by leaf extracts of Cannabis sativa and Morus nigra in liver homogenates such as 230.63±0.52 and 326.91±0.01 µg/mL (iron-induced) while 300.47±0.07 and 300.47±0.07 µg/mL (nitroprusside induced), respectively. The extracts of Cannabis sativa extract showed promising activity (96.04±0.060%) against DPPH radicals while Morus nigra showed a moderate activity (34.11±0.120%). The results suggest that different accessions ofCannabis sativa and Morus nigra are a potential source of antioxidants and have a therapeutic effect against disease induced by oxidative stress and hence can be used for novel drug discovery and development.

Resumo O presente estudo teve como objetivo avaliar o potencial antioxidante e o efeito inibitório de Cannabis sativa e Morus nigra contra a peroxidação lipídica em homogenatos de cérebro e fígado de cabras. A formação de radicais livres, espécies altamente reativas de oxigênio (ROS) e espécies reativas de nitrogênio (RNS), é um processo metabólico normal para sinalização celular e combate aos antígenos. No entanto, eles podem causar sérios danos se forem produzidos em portagens ampliadas. Além disso, distúrbios metabólicos também servem como fontes dessas espécies reativas, embora o problema possa ser resolvido por meio de suplementos e outros fitoquímicos. Neste estudo, duas espécies de plantas foram avaliadas quanto ao seu potencial biológico, empregando um espectro de ensaios antioxidantes. A atividade antioxidante foi realizada por ensaio de peroxidação lipídica. O extrato de água preparado a partir de folhas de Cannabis sativa e Morus nigra mostrou inibição significativa (P < 0,05) em comparação com o controle, ou seja, 522,6 ± 0,06 e 659,97 ± 0,03 µg / mL contra peroxidação lipídica induzida por ferro em homogenato de cérebro de cabra, enquanto as inibições foram 273,54 ± 0,04 e 309,18 ± 0,05 µg / mL contra a peroxidação lipídica do cérebro induzida por nitroprussiato. A peroxidação lipídica induzida por ferro e nitroprussiato também foi significativamente inibida por extratos de folhas de Cannabis sativa e Morus nigra em homogenatos de fígado, como 230,63 ± 0,52 e 326,91 ± 0,01 µg / mL (induzida por ferro), enquanto 300,47 ± 0,07 e 300,47 ± 0,07 µg / mL (induzida por nitroprussiato), respectivamente. Os extratos do extrato de Cannabis sativa apresentaram atividade promissora (96,04 ± 0,060%) contra os radicais DPPH enquanto Morus nigra apresentou atividade moderada (34,11 ± 0,120%). Os resultados sugerem que diferentes acessos de Cannabis sativa e Morus nigra são uma fonte potencial de antioxidantes e têm efeito terapêutico contra doenças induzidas por estresse oxidativo e, portanto, podem ser usados ​​para a descoberta e desenvolvimento de novos medicamentos.

Feral dogs in Chitral gol national park, Pakistan: a potential threat to the future of threatened Kashmir Markhor (Capra falconeri cashmiriensis) / Cães ferozes no parque nacional Chitral gol, Paquistão: uma ameaça potencial para o futuro do Markhor da Caxemira ameaçado (Capra falconeri cashmiriensis)

Abstract Feral dogs are well-organized hunters of ungulates in many parts of the world, causing great damage to wildlife populations and ultimately to the ecosystem. In Pakistan, the impacts of feral dogs on the wildlife have not been documented yet. In a period of fifteen years (2006-2020), feral dogs have killed hundreds of threatened markhor in Chitral gol national park (CGNP), Pakistan. Despite direct predation other impacts including disturbance and competition with other natural predators, could compromise conservation and management efforts. The population of feral dogs seems to have been increased with the increase of dumping sites by communities. Our findings suggest that there are pressing needs of controlling the feral dogs population and eradicating them from the core zone of CGNP and surrounding buffer communities. Conventional culling of dogs should be coupled with modern techniques like castration and sterilization. Communities should be educated regarding the clean environment, proper disposal of home wastes and, biodiversity conservation.

Resumo Os cães ferozes são caçadores bem organizados de ungulados em muitas partes do mundo, causando grandes danos à população de animais selvagens e, em última instância, ao ecossistema. No Paquistão, os impactos dos cães selvagens na vida selvagem ainda não foram documentados. Em um período de 15 anos (2006-2020), cães selvagens mataram centenas de markhor ameaçados no parque nacional Chitral gol (CGNP), Paquistão. Apesar da predação direta, outros impactos, incluindo distúrbios e competições com outros predadores naturais podem comprometer os esforços de conservação e manejo. A população de cães selvagens parece ter aumentado com o aumento dos locais de despejo pelas comunidades. Nossas descobertas sugerem que há necessidades urgentes para controlar a população de cães selvagens e erradicá-los da zona central do CGNP e das comunidades-tampão vizinhas. O abate convencional de cães deve ser combinado com técnicas modernas como castração e esterilização. As comunidades devem ser educadas sobre o meio ambiente limpo, o descarte adequado de resíduos domésticos e a conservação da biodiversidade.

Preparation of colloidal gold test strips for the detection of antibodies to peste des petits ruminants based on monoclonal antibodies to N protein / 生物工程学报

A simple, fast, and visual method for detecting antibodies against peste des petits ruminants virus (PPRV) using colloidal gold strips was developed. In this study, the pET-32a-N was transformed into Escherichia coli Rosetta (DE3) for expression. Hybridoma cell lines were generated by fusing SP2/0 myeloma cells with splenocytes from immunized mice with the expressed and purified N protein of PPRV. The PPRV N protein was labeled with colloidal gold particles as the gold-labeled antigen. The N protein served as the gold standard antigen and as the test (T) line-coated antigen, while the monoclonal antibody served as the quality control (C) line-coated antibody to assemble the colloidal gold immunochromatographic test strips for detecting antibodies against the N protein of PPRV. Hybridoma cell line designated as 1F1 was able to stably secrete the monoclonal antibody against the N protein of PPRV. The titer of 1F1 monoclonal antibody in ascites was 1:128 000 determined by indirect enzyme-linked immunosorbent assays (ELISA), and the immunoglobulin subtype of the monoclonal antibody was IgG1, with kappa chain. The obtained monoclonal antibody was able to specifically recognize the N protein of PPRV, as shown by Western blotting and indirect immunofluorescent assay (IFA). The developed colloidal gold test strip method was able to detect PPRV antibodies specifically, and there was no difference between different batches of the test strips. Testing of a total of 122 clinical sera showed that the compliance rate of the test strip with ELISA test was 97.6%.The test strip assay developed in this study has good specificity, reproducibility, and sensitivity, and it can be used for the rapid detection of PPRV antibodies.

BLOC1S1 promotes proliferation of goat spermatogonial stem cells / 生物工程学报

With the rapid development of gene editing technology, the study of spermatogonial stem cells (SSCs) holds great significance in understanding spermatogenesis and its regulatory mechanism, developing transgenic animals, gene therapy, infertility treatment and protecting rare species. Biogenesis of lysosome-related organelles complex 1 subunit 1 (BLOC1S1) is believed to have anti-brucella potential. Exploring the impack of BLOC1S1 on goat SSCs not only helps investigate the ability of BLOC1S1 to promote SSCs proliferation, but also provides a cytological basis for disease-resistant breeding research. In this study, a BLOC1S1 overexpression vector was constructed by homologous recombination. The BLOC1S1 overexpression cell line of goat spermatogonial stem cells was successfully constructed by lentivirus packaging, transfection and puromycin screening. The overexpression efficiency of BLOC1S1 was found to be 18 times higher using real time quantitative PCR (RT-qPCR). Furthermore, the results from cell growth curve analysis, flow cytometry for cell cycle detection, and 5-ethynyl-2'-deoxyuridine (EdU) staining showed that BLOC1S1 significantly increased the proliferation activity of goat SSCs. The results of RT-qPCR, immunofluorescence staining and Western blotting analyses revealed up-regulation of proliferation-related genes (PCNA, CDK2, CCND1), and EIF2S3Y, a key gene regulating the proliferation of spermatogonial stem cells. These findings strongly suggest that the proliferative ability of goat SSCs can be enhanced through the EIF2S3Y/ERK pathway. In summary, this study successfully created a goat spermatogonial stem cell BLOC1S1 overexpression cell line, which exhibited improved proliferation ability. This research laid the groundwork for exploring the regulatory role of BLOC1S1 in goat spermatogonia and provided a cell platform for further study into the biological function of BLOC1S1. These findings also establish a foundation for breeding BLOC1S1 overexpressing goats.

miR-23b-3p regulates the differentiation of goat intramuscular preadipocytes by targeting the PDE4B gene / 生物工程学报

This study aimed to explore the effect of miR-23b-3p on the differentiation of goat intramuscular preadipocytes, and to confirm whether miR-23b-3p plays its roles via targeting the PDE4B gene. Based on the pre-transcriptome sequencing data obtained previously, the miR-23b-3p, which was differentially expressed in goat intramuscular adipocytes before and after differentiation, was used as an entry point. real-time quantitative-polymerase chain reaction (qPCR) was used to detect the expression pattern of miR-23b-3p during the differentiation of goat intramuscular preadipocytes. The effects of miR-23b-3p on adipose differentiation and adipose differentiation marker genes were determined at the morphological and molecular levels. The downstream target genes of miR-23b-3p were determined using bioinformatics prediction as well as dual luciferase reporter assay to clarify the targeting relationship between miR-23b-3p and the predicted target genes. The results indicated that overexpression of miR-23b-3p reduced lipid droplet accumulation in goat intramuscular adipocytes, significantly down-regulated the expression levels of adipogenic marker genes AP2, C/EBPα, FASN, and LPL (P < 0.01). In addition, the expressions of C/EBPβ, DGAT2, GLUT4 and PPARγ were significantly downregulated (P < 0.05). After interfering with the expression of miR-23b-3p, lipid droplet accumulation was increased in goat intramuscular adipocytes. The expression levels of ACC, ATGL, AP2, DGAT2, GLUT4, FASN and SREBP1 were extremely significantly up-regulated (P < 0.01), and the expression levels of C/EBPβ, LPL and PPARγ were significantly up-regulated (P < 0.05). It was predicted that PDE4B might be a target gene of miR-23b-3p. The mRNA expression level of PDE4B was significantly decreased after overexpression of miR-23b-3p (P < 0.01), and the interference with miR-23b-3p significantly increased the mRNA level of PDE4B (P < 0.05). The dual luciferase reporter assay indicated that miR-23b-3p had a targeting relationship with PDE4B gene. MiR-23b-3p regulates the differentiation of goat intramuscular preadipocytes by targeting the PDE4B gene.

Quantification of antigen of Mycoplasma capricolum subsp. capripneumoniae by optical assay / 生物工程学报

Mycoplasma capricolum subsp. capripneumoniae (Mccp) is the cause of contagious caprine pleuropneumonia (CCPP) in goats. Inactivated vaccines and capsular polysaccharide (CPS) indirect hemagglutination reagents are available for prevention and serological detection, but high culture costs and complex antigen quantification have been plagued by production staff. In order to solve these problems in production practice, a sugar fermentation medium with an initial pH value of 7.8, which could improve the production of two antigens simultaneously, was screened out by changing the initial pH value based on previous Mccp metabolomics analysis. Since phenol red can be identified by UV absorption spectrum and cetyltrimethylammonium bromide (CTAB) can bind to anionic capsular polysaccharide, a UV spectrum measurement method for analyzing the culture stage reached by Mccp and a CTAB precipitation test for relative quantification of capsular polysaccharide antigen content in the fermentation broth were established. The UV spectrum observation method can guide the production of Mccp according to the growth curve of Mccp, which greatly reduces the monitoring time of the traditional CCU method and improves the accuracy of the original eye-observation method. The established CTAB precipitation test can complete the monitoring of CPS content within 5 hours, which greatly reduces the time required compared with the traditional differential technique, and its accuracy was verified by the phenol-sulfuric acid method. The optimized culture medium and the two correlation comparison methods established in this study can effectively reduce the production cost of Mccp and improve the production efficiency. The two assays have been used in the research at our laboratory, which provides experimental data for further improvement of the production process of CCPP inactivated vaccine and capsular polysaccharide as well as rapid quantification.

The inhibitory effect of Cannabis Sativa L. and Morus nigra L. against lipid peroxidation in goat liver and brain homogenates / O efeito inibitório de Cannabis Sativa L. e Morus nigra L. contra a peroxidação lipídica em homogenatos de fígado e cérebro de cabra

The present study was aimed to evaluate the antioxidant potential and inhibitory effect of Cannabis sativa and Morus nigra against lipid peroxidation in goat brain and liver homogenates. The formation of free radicals, highly reactive oxygen species (ROS) and reactive nitrogen species (RNS) is a normal metabolic process for cellular signaling and countering the antigens. However, they may cause serious damage if they produced at amplified tolls. In addition, metabolic disorders also serve as sources of these reactive species. Although the issue can be addressed through supplements and other phytochemicals. In this study, two plant species were evaluated for their biological potential by employing a spectrum of antioxidant assays. The antioxidant activity was performed by lipid peroxidation assay. The water extract prepared from leaves of Cannabis sativa and Morus nigra showed significant (P<0.05) inhibition as compared to control i.e., 522.6±0.06 and 659.97±0.03 µg/mL against iron-induced lipid peroxidation in goat brain homogenate while the inhibitions were 273.54±0.04 and 309.18±0.05 µg/mL against nitroprusside induced lipid peroxidation of the brain. The iron and nitroprusside induced lipid peroxidation was also significantly inhibited by leaf extracts of Cannabis sativa and Morus nigra in liver homogenates such as 230.63±0.52 and 326.91±0.01 µg/mL (iron-induced) while 300.47±0.07 and 300.47±0.07 µg/mL (nitroprusside induced), respectively. The extracts of Cannabis sativa extract showed promising activity (96.04±0.060%) against DPPH radicals while Morus nigra showed a moderate activity (34.11±0.120%). The results suggest that different accessions of Cannabis sativa and Morus nigra are a potential source of antioxidants and have a therapeutic effect against disease induced by oxidative stress and hence can be used for novel drug discovery and development.

O presente estudo teve como objetivo avaliar o potencial antioxidante e o efeito inibitório de Cannabis sativa e Morus nigra contra a peroxidação lipídica em homogenatos de cérebro e fígado de cabras. A formação de radicais livres, espécies altamente reativas de oxigênio (ROS) e espécies reativas de nitrogênio (RNS), é um processo metabólico normal para sinalização celular e combate aos antígenos. No entanto, eles podem causar sérios danos se forem produzidos em portagens ampliadas. Além disso, distúrbios metabólicos também servem como fontes dessas espécies reativas, embora o problema possa ser resolvido por meio de suplementos e outros fitoquímicos. Neste estudo, duas espécies de plantas foram avaliadas quanto ao seu potencial biológico, empregando um espectro de ensaios antioxidantes. A atividade antioxidante foi realizada por ensaio de peroxidação lipídica. O extrato de água preparado a partir de folhas de Cannabis sativa e Morus nigra mostrou inibição significativa (P < 0,05) em comparação com o controle, ou seja, 522,6 ± 0,06 e 659,97 ± 0,03 µg / mL contra peroxidação lipídica induzida por ferro em homogenato de cérebro de cabra, enquanto as inibições foram 273,54 ± 0,04 e 309,18 ± 0,05 µg / mL contra a peroxidação lipídica do cérebro induzida por nitroprussiato. A peroxidação lipídica induzida por ferro e nitroprussiato também foi significativamente inibida por extratos de folhas de Cannabis sativa e Morus nigra em homogenatos de fígado, como 230,63 ± 0,52 e 326,91 ± 0,01 µg / mL (induzida por ferro), enquanto 300,47 ± 0,07 e 300,47 ± 0,07 µg / mL (induzida por nitroprussiato), respectivamente. Os extratos do extrato de Cannabis sativa apresentaram atividade promissora (96,04 ± 0,060%) contra os radicais DPPH enquanto Morus nigra apresentou atividade moderada (34,11 ± 0,120%). Os resultados sugerem que diferentes acessos de Cannabis sativa e Morus nigra são uma fonte potencial de antioxidantes e têm efeito terapêutico [...].

Gene cloning and sequence analysis of the RPL29 gene and its effect on lipogenesis in goat intramuscular adipocytes / 生物工程学报

The aim of this study was to clone the goat RPL29 gene and analyze its effect on lipogenesis in intramuscular adipocytes. Using Jianzhou big-eared goats as the object, the goat RPL29 gene was cloned by reverse transcription-polymerase chain reaction (RT-PCR), the gene structure and expressed protein sequence were analyzed by bioinformatics, and the mRNA expression levels of RPL29 in various tissues and different differentiation stages of intramuscular adipocytes of goats were detected by quantitative real-time PCR (qRT-PCR). The RPL29 overexpression vector pEGFP-N1-RPL29 constructed by gene recombination was used to transfect into goat intramuscular preadipocytes and induce differentiation. Subsequently, the effect of overexpression of RPL29 on fat droplet accumulation was revealed morphologically by oil red O and Bodipy staining, and changes in the expression levels of genes related to lipid metabolism were detected by qRT-PCR. The results showed that the length of the goat RPL29 was 507 bp, including a coding sequence (CDS) region of 471 bp which encodes 156 amino acid residues. It is a positively charged and stable hydrophilic protein mainly distributed in the nucleus of cells. Tissue expression profiling showed that the expression level of this gene was much higher in subcutaneous adipose tissue and inter-abdominal adipose tissue of goats than in other tissues (P < 0.05). The temporal expression profile showed that the gene was expressed at the highest level at 84 h of differentiation in goat intramuscular adipocytes, which was highly significantly higher than that in the undifferentiated period (P < 0.01). Overexpression of RPL29 promoted lipid accumulation in intramuscular adipocytes, and the optical density values of oil red O staining were significantly increased (P < 0.05). In addition, overexpression of RPL29 was followed by a highly significant increase in ATGL and ACC gene expression (P < 0.01) and a significant increase in FASN gene expression (P < 0.05). In conclusion, the goat RPL29 may promote intra-muscular adipocyte deposition in goats by up-regulating FASN, ACC and ATGL.

Cloning and expression profile of ZFP36L1 gene in goat / 生物工程学报

The purpose of this study was to clone and characterize the ZFP36L1 (zinc finger protein 36-like 1) gene, clarify its expression characteristics, and elucidate its expression patterns in different tissues of goats. Samples of 15 tissues from Jianzhou big-eared goats, including heart, liver, spleen, lung and kidney were collected. Goat ZFP36L1 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR), then the gene and protein sequence were analyzed by online tools. Quantitative real-time polymerase chain reaction (qPCR) was used to detect the expression level of ZFP36L1 in intramuscular preadipocytes in different tissues and adipocytes of goat at different differentiation stages. The results showed that the length of ZFR36L1 gene was 1 224 bp, and the coding sequence (CDS) region was 1 017 bp, encoding 338 amino acids, which was a non-secretory unstable protein mainly located in nucleus and cytoplasm. Tissue expression profile showed that ZFP36L1 gene was expressed in all selected tissues. In visceral tissues, the small intestine showed the highest expression level (P < 0.01). In muscle tissue, the highest expression level was presented in longissimus dorsi muscle (P < 0.01), whereas the expression level in subcutaneous adipose tissue was significantly higher than that in other tissues (P < 0.01). The results of induced differentiation showed that the expression of this gene was up-regulated during adipogenic differentiation of intramuscular precursor adipocytes (P < 0.01). These data may help to clarify the biological function of the ZFP36L1 gene in goat.

Cloning, identification and functional analysis of the goat transcription factor c-fos / 生物工程学报

C-fos is a transcription factor that plays an important role in cell proliferation, differentiation and tumor formation. The aim of this study was to clone the goat c-fos gene, clarify its biological characteristics, and further reveal its regulatory role in the differentiation of goat subcutaneous adipocytes. We cloned the c-fos gene from subcutaneous adipose tissue of Jianzhou big-eared goats by reverse transcription-polymerase chain reaction (RT-PCR) and analyzed its biological characteristics. Using real-time quantitative PCR (qPCR), we detected the expression of c-fos gene in the heart, liver, spleen, lung, kidney, subcutaneous fat, longissimus dorsi and subcutaneous adipocytes of goat upon induced differentiation for 0 h to 120 h. The goat overexpression vector pEGFP-c-fos was constructed and transfected into the subcutaneous preadipocytes for induced differentiation. The morphological changes of lipid droplet accumulation were observed by oil red O staining and bodipy staining. Furthermore, qPCR was used to test the relative mRNA level of the c-fos overexpression on adipogenic differentiation marker genes. The results showed that the cloned goat c-fos gene was 1 477 bp in length, in which the coding sequence was 1 143 bp, encoding a protein of 380 amino acids. Protein structure analysis showed that goat FOS protein has a basic leucine zipper structure, and subcellular localization prediction suggested that it was mainly distributed in the nucleus. The relative expression level of c-fos was higher in the subcutaneous adipose tissue of goats (P < 0.05), and the expression level of c-fos was significantly increased upon induced differentiation of subcutaneous preadipocyte for 48 h (P < 0.01). Overexpression of c-fos significantly inhibited the lipid droplets formation in goat subcutaneous adipocytes, significantly decreased the relative expression levels of the AP2 and C/EBPβ lipogenic marker genes (P < 0.01). Moreover, AP2 and C/EBPβ promoter are predicted to have multiple binding sites. In conclusion, the results indicated that c-fos gene was a negative regulatory factor of subcutaneous adipocyte differentiation in goats, and it might regulate the expression of AP2 and C/EBPβ gene expression.

Comparison of two luminescence detection methods for staphylococcal enterotoxin C content in simulated milk samples / 细胞与分子免疫学杂志

Objective To compare the sensitivity and accuracy of amplified luminescent proximity homogeneous assay linked immunosorbent assay (AlphaLISA) and magnetic particles-based chemiluminescence immunoassay (MP-CLIA) for detection of staphylococcal enterotoxin C (SEC) in the simulated milk samples. Methods The AlphaLISA was constructed using goat anti-SEC polyclonal antibody-coupled receptor microspheres, biotin-labeled SEC monoclonal antibody and streptavidin-coupled donor microspheres. The MP-CLIA was constructed using goat anti-SEC polyclonal antibody conjugated alkaline phosphatase, biotin-labeled anti-SEC monoclonal antibody and streptavidin conjugated magnetic beads. Results The sensitivity of AlphaLISA to detect SEC content in simulated milk samples was 4.04 ng/L, and the coefficient of variation (CV) was 1.98%~9.82%. The sensitivity of MP-CLIA was 108.19 ng/L and CV was 4.63%~20.40%. Conclusion Compared with MP-CLIA, AlphaLISA is more sensitive and accurate to detecting SEC.

Padronização de uma PCR multiplex para a detecção de fraude em leite bubalino e caprino por adição de leite bovino / Standardization of a multiplex PCR for detection of fraud in buffalo and goat milk by addition of bovine milk

O consumo de leite de espécies como bubalino e caprino tem se popularizado por representarem uma alternativa para indivíduos que possuem restrições alimentares relacionadas ao leite bovino e em virtude das propriedades nutricionais desses alimentos. No entanto, fatores como a baixa produção e a sazonalidade predispõem a adulterações destes alimentos, principalmente pela adição de leite bovino, visando maior rendimento e lucratividade. Assim, o objetivo do estudo foi padronizar um método de PCR multiplex para autenticação de leites bubalino e caprino. Para isso, amostras de leite exclusivamente de cada espécie foram utilizados para a padronização da técnica. Em seguida, foi realizada a fraude pela adição de leite bovino ao caprino e ao bubalino, em proporções de 0,1% até 100%. A técnica foi eficaz, precisa, rápida e prática para a detecção do DNA de bovino, bubalino e caprino, separadamente e em conjunto. Na fraude experimental, o limite de detecção da técnica ocorreu a partir do menor percentual testado (0,1%) tanto no leite caprino quanto no bubalino. Dessa forma, a PCR multiplex testada mostrou ser uma importante ferramenta para a autenticação de leite, pendendo ser utilizada para fins de fiscalização por órgãos competentes.

Milk consumption of species such as buffalo and goat has become popular due to the nutritional properties of these foods and because they represent an alternative for individuals who have dietary restrictions related to bovine milk. However, factors such as low production and seasonality predispose to adulteration, mainly by the addition of bovine milk, aiming at higher yield and profitability. Thus, the aim of the present study was to standard a multiplex PCR method for buffalo and goat milks authentication. For this, the milks exclusively of each species were used to standardize the technique. Subsequently, fraud was performed by the addition of bovine milk to goat and buffalo in proportions from 0.1% to 100%. The technique was effective and accurate for detecting bovine, buffalo and goat DNA separately and together quickly and practically. In experimental fraud, the detection limit of the technique occurred from the lowest percentage tested (0.1%) in both goat and buffalo milk. Thus, the multiplex PCR tested proved to be an important tool for milk authentication, pending to be used for supervision by competent agencies.

Potencial antioxidante de hidrolisados proteicos obtidos a partir de caseinato de ruminantes pela ação de proteases comerciais / Potential antioxidant of protein hydrolysates obtained from ruminant caseinate by the action of commercial proteases

Introdução: a oxidação em sistemas biológicos está relacionada ao desenvolvimento de patologias em humanos. A ingestão de alimentos ricos em compostos químicos que exercem atividade antioxidante contribui para a prevenção e redução dos efeitos deletérios dos radicais livres formados no organismo. Peptídeos derivados das caseínas têm mostrado um elevado potencial como agentes antioxidantes. Objetivos: neste sentido, o presente estudo avaliou a atividade antioxidante de hidrolisados derivados de caseínas de leites das espécies bubalina, bovina e caprina, obtidos pela ação de diferentes proteases. Metodologia: inicialmente, as caseínas foram isoladas dos demais componentes do leite, depois foram submetidas ao processo de proteólise pelas enzimas bromelina, papaína, tripsina e neutrase, individualmente. A atividade antioxidante dos hidrolisados foi avaliada, através da capacidade de eliminação dos radicais: hidroxila (OH­Ë™), superóxido (O2­Ë™), 2,2 difenil-1-picrilhidrazil (DPPH˙), 2,2'azinobis-(3-ácido etilbenzotiazolino-6-sulfônico (ABTS˙), e quelante dos íons metálicos cobre (Cu2+) e ferro (Fe2+). Resultados: os resultados mostraram que a caseína bovina apresentou o menor (35,54%) grau de hidrólise e a caseína bubalina apresentou o maior (85,64%) grau de hidrólise pela ação da neutrase e bromelina após 480 minutos, respectivamente. O potencial para o sequestro dos radicais hidroxila variou entre 0 e 100%, superóxido superior a 80%, ABTS superior a 85%, DPPH entre 20 e 95% habilidade de quelar ferro entre 10 e 100% e cobre entre 14 e 80%. Conclusão: assim, a hidrólise das caseínas do leite bubalino, bovino e caprino foram capazes de produzir hidrolisados com elevado potencial antioxidante e que, mediante novos estudos, poderá vir ser incorporado em produtos alimentícios para o consumo humano.

Introduction: oxidation in biological systems is related to the development of pathologies in humans. The ingestion of foods rich in chemical compounds that exert antioxidant activity contributes to the prevention and reduction of the deleterious effects of free radicals formed in the body. Peptides derived from caseins have shown high potential as antioxidant agents. Objectives: the present study evaluated the antioxidant activity of casein hydrolysates derived from bubaline, bovine, and caprine milk obtained by the action of different proteases. Methodology: initially, the caseins were isolated from the other milk components, and then subjected to the proteolysis process by the enzymes bromelain, papain, trypsin and neutrase, individually. The antioxidant activity of the hydrolysates was evaluated, through the capacity of elimination of the radicals: hydroxyl (OH-˙), superoxide (O2-˙), 2,2 diphenyl-1-picrylhydrazyl (DPPH˙), 2,2'azinobis-(3-ethylbenzothiazolino-6-sulfonic acid (ABTS˙), and chelating of the metal ions copper (Cu2+) and iron (Fe2+). Results: the results showed that bovine casein showed the lowest (35.54%) degree of hydrolysis and bubaline casein showed the highest (85.64%) degree of hydrolysis by the action of neutrase and bromelin after 480 minutes, respectively. The potential for hydroxyl radical sequestration varied between 0 and 100%, superoxide higher than 80%, ABTS higher than 85%, DPPH between 20 and 95% and the ability to chelate iron between 10 and 100% and copper between 14 and 80%. Conclusion: thus, the hydrolysis of caseins from bubaline, bovine and goat milk were able to produce hydrolysates with high antioxidant potential and that, upon further studies, may be incorporated into food products for human consumption.

Diagnosis of goat tuberculosis using tuberculinization and molecular techniques

Tuberculosis is an infectious, chronic, and worldwide disease. It has been known since the beginning of humanity and still negatively influences public health and livestock, especially, in Brazil, in the northeast. Etiologic agents are the mycobacteria of the Mycobacterium tuberculosis complex, which is the most important in mammals' involvement. The state of Bahia has 68.7% of its territory located in the semiarid region and holds the largest goat herd in the country. Goat breeding is a social and economic activity that adds value to this region. Up to the present, data on goat tuberculosis is unknown in this state. Thus, this study seeks data on tuberculosis prevalence in goats in a semiarid region of Bahia by using the comparative tuberculin test and multiplex polymerase chain reaction (PCR). A total of 600 adult animals of both sexes were evaluated. A prevalence of 0.33% (2/600) and 33.33% (1/3) properties were found for positive animals. Each assessed property had a questionnaire to analyze the epidemiological data management and relevant aspects for the disease occurrence. To confirm the positive tuberculin test results, PCR was used to detect and identify the pathogenic mycobacteria involved in the infection. It is concluded that most of the properties performing goat breeding in the region show low technification levels and promote farming between different species. Low prevalence of the disease alerts preventive measures to avoid major proportion situations that could influence the goat breeding in the state.

Anatomical and morphometric study of goat middle ear ossicles (Capra aegagrus hircus) / [Estudo anatômico e morfométrico dos ossículos da orelha média de caprinos (Capra aegagrus hircus)]

The social and economic roles of goat farming in Northeastern Brazil, allied to the fact that the use of goat middle ear ossicles for research and human ear surgery training has not yet been proposed, justify the study of their applicability as an experimental model. The middle ears of 19 goats (Capra aegagrus hircus) from the bone collection of the Laboratory and Didactic Anatomy Museum of Domestic and Wild Animals of the Federal University of Vale do São Francisco (UNIVASF) were dissected. The malleus, incus, and stapes were evaluated regarding their macroscopic morphology and biometry (length, width, and height). Ossicle morphology was similar to sheep, human, and bovine morphology. The malleus was 1.3 times heavier and 2.2 times longer than the incus, and 9.0 times heavier and 3.7 times longer than the stapes. The size relationship was positive between the stapes and the malleus and negative between the stapes and the incus. It is concluded that the middle ear size and the anatomical similarities with human ossicles make goats a useful model for experimental scientific studies, reconstructive surgery practice of the ossicular chain, and human ear surgery training.(AU)

Tanto o papel social quanto o econômico da caprinocultura na região Nordeste do Brasil, somados ao fato de que o uso de ossículos da orelha média de caprinos para estudos e treinamento cirúrgico otológico humano ainda não foi proposto, justificam o estudo de sua aplicabilidade como modelo experimental. Foram dissecadas as orelhas médias de 19 caprinos (Capra aegagrus hircus), provenientes do ossuário do Laboratório e Museu Didático de Anatomia dos Animais Domésticos e Silvestres - Universidade Federal do Vale do São Francisco. Martelos, bigornas e estribos tiveram a morfologia macroscópica e a biometria (comprimento, largura e altura) avaliadas. A morfologia dos ossículos assemelhou-se a de ovinos, humanos e bovinos. O martelo foi 1,3 vez mais pesado e 2,2 vezes mais comprido que a bigorna e 9,0 vezes mais pesado e 3,7 vezes mais comprido que o estribo. A relação de tamanho entre o estribo e o martelo foi positiva, e entre o estribo e a bigorna negativa. Conclui-se que o tamanho da orelha média e as semelhanças anatômicas com os ossículos humanos tornam os caprinos um modelo útil para estudos científicos experimentais, prática cirúrgica reconstrutiva da cadeia ossicular e treinamento cirúrgico otológico humano.(AU)

Dairy goat production in the semi-arid region: productive and reproductive analysis, and the influence of the adoption of hygienic practices on milk quality / Produção de cabras leiteiras no semiárido: análise produtiva e reprodutiva, bem como influência da adoção de práticas higiênicas na qualidade do leite

The aim of this study was to evaluate the productive and reproductive performance of dairy goat genotypes, as well as the influence of hygienic practices by farmers on the quality of goat milk. Productive and reproductive data of 81 Anglo Nubian and Alpine crossbred goats, as well as data on total milk production of goats over 100 days of lactation and partial production up to 56 days of lactation. Also, in natura milk samples from 160 properties in the region were evaluated. At the time of collection, a questionnaire was applied to diagnose goat milk production systems. Data from productive and reproductive performance were evaluated by Tukey test and descriptive analysis using the SPSS program. The herd presented a variation of the total milk production in lactation (TMP) from 267.40kg to 468.55kg, with lactation length ranging from 157 to 247 days, and average daily production between 1.43 and 1.89kg/day. Fertility rates were satisfactory, with the lowest rate being 76% and the highest 92%, with an average of 85.24% considering the six seasons of birth. The means of the gestation periods varied between 144 and 152 days. Regarding the sanitary characterization, 73% of farmers performed a cleaning of the room before and after milking. However, 94.8% of farmers did not eliminate the first jets of milk and only 29.2% used the screened mug test to identify clinical mastitis. Only 41% of farmers performed pre and post-dipping and 30.2% applied the iodine solution. Only 8.3% of farmers used disposable paper towels. However, 92% of producers still used fabric towel. It was also observed that 99% of the properties stored milk in buckets or cans without refrigeration. In the microbiological analysis, a small amount of milk samples (5.6%) was contaminated with Staphylococcus aureus, however the total coliform count was high. Regarding the somatic cell count, it was found that 86% of the properties presented values above one million cells per mL of sample. The study demonstrated the prevalence of several factors that contribute to the vulnerability of milk contamination in various stages of production such as milking and processing. Thus, the guidance and awareness of those responsible is extremely important to improve goat milk quality in the semi-arid region of Paraíba.(AU)

O objetivo do presente estudo foi avaliar o desempenho produtivo e reprodutivo de genótipos caprinos leiteiros, assim como a influência da adoção das práticas higiênicas pelos produtores sobre a qualidade do leite caprino. A primeira parte da pesquisa foi realizada na Estação Experimental Pendência-PB. Foram analisados dados produtivos e reprodutivos de 81 cabras Anglo-Nubiana e alpinas mestiças, bem como dados de produção total de leite das cabras acima de 100 dias de lactação e produção parcial até 56 dias de lactação. Também foram avaliadas amostras de leite in natura de 160 propriedades da região. No momento da coleta, foi aplicado um questionário de diagnóstico dos sistemas de produção de leite caprino. Os dados de desempenho produtivo e reprodutivo foram avaliados pelo teste de Tukey e pela análise descritiva utilizando o programa SPSS. O rebanho apresentou uma variação da produção de leite total na lactação desde 267,40kg a 468,55kg, com durações de lactação variando de 157 e 247 dias, e produção média diária entre 1,43 e 1,89kg/dia. As taxas de fertilidade foram satisfatórias, sendo a menor taxa de 76% e a maior de 92%, com uma média de 85,24% considerando as seis estações de parição. As médias dos períodos de gestação variaram entre 144 e 152 dias. Quanto à caracterização da sanidade, 73% dos produtores realizam limpeza da sala antes e após a ordenha. No entanto, 94,8% dos produtores não eliminam os primeiros jatos de leite e somente 29,2% utilizam o teste da caneca telada para identificação de mastite clínica. Apenas 41% dos produtores realizam pré-dipping e pós-dipping e 30,2% aplicam a solução de iodo. Somente 8,3% dos produtores usam toalhas descartáveis e 92% ainda utilizam toalha de tecido. Observou-se, ainda, que 99% das propriedades armazenam o leite ordenhado em baldes ou latões, sem refrigeração. Pequenas quantidades de amostras (5,6%) estavam contaminadas por Staphylococcus aureus, porém a contagem de coliformes totais teve valor elevado. Em relação à contagem de células somáticas, verificou-se que 86% das propriedades apresentaram CCS acima de um milhão de células por mL, devendo-se ajustar corretamente os manejos alimentar, produtivo e reprodutivo. As épocas de parição, lactação e o genótipo influenciaram de forma direta os índices produtivos e reprodutivos dos animais. Obsevou-se a prevalência de vários fatores que contribuem para a vulnerabilidade de contaminação do leite em diversas etapas de produção, tais como ordenha e processamento. Portanto, a orientação adequada e a conscientização dos responsáveis são de extrema importância para melhorar a qualidade do leite de cabra na região semiárida da Paraíba.(AU)

Intake, digestibility, ingestive behavior and performance of goats fed spineless cactus genotypes resistant to carmine cochineal / Consumo, digestibilidade, comportamento ingestivo e desempenho de caprinos alimentados com genótipos de palma forrageira resistentes à cochonilha-do-carmim

The objective was to evaluate the intake and digestibility of nutrients, ingestive behavior, and performance of goats fed with spineless cactus genotypes resistant to carmine cochineal (Miúda or Orelha de Elefante Mexicana (OEM). Thirty castrated male goats, without defined breed, aged 12 to 14 months, with an average body weight of 19.0±2.8kg, were distributed in a completely randomized design among three treatments (Control - Tifton hay, Miúda, and OEM) and ten replicates; the initial weight was considered as the covariate. The intake of organic matter (OM), neutral detergent fiber (NDF) was highest in the control treatment, while the intake of NFC was higher in the OEM diet. Treatments containing forage cactus showed the highest digestibility of DM, OM, and NFC. The animals fed the control diet spent more time on rumination and total chewing, but the time spent feeding or feeding efficiency, and performance did not differ. The use of spineless cactus genotypes resistant to carmine cochineal (Miúda and Orelha de Elefante Mexicana) in a diet for goats, in the amount of 450g/kg of DM does not interfere with the performance of the animals and improves the digestibility of OM and NFC.(AU)

Objetivou-se com o presente estudo avaliar o consumo e a digestibilidade de nutrientes, o comportamento ingestivo e o desempenho de caprinos alimentados com genótipos de palma forrageira resistentes à cochonilha-carmim (miúda ou orelha-de-elefante-mexicana (OEM). Trinta cabritos machos, castrados, sem raça definida, com idades entre 12 e 14 meses e peso corporal médio de 19,0±2,8kg, foram distribuídos em delineamento inteiramente ao acaso, em três tratamentos (controle - feno de tifton; miúda e OEM) e 10 repetições; o peso inicial foi considerado a covariável. O consumo de matéria orgânica (MO) e de fibra em detergente neutro (FDN) foi maior no tratamento controle; enquanto a ingestão de CNF foi maior na dieta OEM. Tratamentos contendo palma forrageira apresentaram as maiores digestibilidades de MS, MO e CNF. Os animais alimentados com a dieta controle gastaram mais tempo em ruminação e em mastigação total, mas o tempo gasto com alimentação ou a eficiência alimentar e o desempenho não diferiram. A utilização de genótipos de palma forrageira resistentes à cochonilha-carmim (miúda e orelha-de-elefante-mexicana) na dieta de caprinos, na quantidade de 450g/kg de MS, não interfere no desempenho dos animais e melhora a digestibilidade de MO e CNF.(AU)