Chromatin condensation but not DNA integrity of pig sperm is greater in the sperm-rich fraction

Background Protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo development.In some mammals,like pigs,ejaculates are emitted in three separate fractions:pre-sperm,sperm-rich(SRF)and post sperm-rich(PSRF).These fractions are known to vary in volume,sperm concentration and quality,as well as in the origin and composition of seminal plasma(SP),with differences being also observed within the SRF one.Yet,whether disparities in the DNA integrity and chromatin condensation and pro-tamination of their sperm exist has not been interrogated.Results This study determined chromatin protamination(Chromomycin A3 test,CMA_(3)),condensation(Dibromobi-mane test,DBB),and DNA integrity(Comet assay)in the pig sperm contained in the first 10 m L of the SRF(SRF-P1),the remaining portion of the sperm-rich fraction(SRF-P2),and the post sperm-rich fraction(PSRF).While chromatin protamination was found to be similar between the different ejaculate fractions(P>0.05),chromatin condensation was seen to be greater in SRF-P1 and SRF-P2 than in the PSRF(P=0.018 and P=0.004,respectively).Regarding DNA integrity,no differences between fractions were observed(P>0.05).As the SRF-P1 has the highest sperm concentra-tion and ejaculate fractions are known to differ in antioxidant composition,the oxidative stress index(OSi)in SP,calcu-lated as total oxidant activity divided by total antioxidant capacity,was tested and confirmed to be higher in the SRF-P1 than in SRF-P2 and PSRF(0.42±0.06 vs.0.23±0.09 and 0.08±0.00,respectively;P<0.01);this index,in addition,was observed to be correlated to the sperm concentration of each fraction(Rs=0.973;P<0.001).Conclusion While sperm DNA integrity was not found to differ between ejaculate fractions,SRF-P1 and SRF-P2 were observed to exhibit greater chromatin condensation than the PSRF.This could be related to the OSi of each fraction.

Sero-Prevalence and Risk factors of Contagious Bovine Pleuropneumonia (CBPP) in Afgoye District Lower Shabelle Region, Somalia

Contagious bovine Pleuropneumonia (CBPP) is an infectious and highly contagious respiratory disease of cattle and water buffalo, caused by the Mycoplasma mycoides subspecies mycoides. It induces significant economic losses and leads to a severe livestock production problem, negatively influencing people’s livelihoods of affected countries. In Somalia, there is no updated data on the prevalence and distribution of the disease. Hence, a descriptive cross-sectional study was conducted from September 2022 to June 2023 in different villages under the Afgoye District of lower Shabelle region, Somalia. The main purpose of this study is to assess the sero-prevalence and identify the associated risk factors for the occurrence of the disease. In this study, villages, age, sex, breed, and body condition were considered as risk factors. A total of 90 blood samples were collected and tested in the laboratory using the Anti-CBPP Elisa kit test. Out of 90 serum samples from herd cattle, 32 were positive, resulting in an overall prevalence of 35.5%. In addition, we found a statistically significant variation between the prevalence of the disease and factors such as sex, age, body condition and breeds. In summary, the overall prevalence of Contagious bovine pleuropneumonia in this study area is worth to be considered because there is a low quality of health care and less awareness of the Contagious bovine Pleuropneumonia effects on herds, which warrants the official authorities to act and follow appropriate preventive and control measures to reduce the incidence of the disease and generate appropriate controlling and prevention measures in all regions of Somalia.

A staggering rise of HIV cases in Pakistan

In the last decade,Pakistan has experienced multidemics of HIV in key populations,namely:injecting drug users,male sex with male,female sex workers,transgender sex workers,and transgenders.According to recent reports,in Pakistan,210000 people with HIV accounts for less than 0.2%of the general population.

Effect of dephytinization on bioavailability of iron,calcium and zinc from infant cereals assessed in the Caco-2 cell model

AIM:To test the effect of the dephytinization of three different commercial infant cereals on iron, calcium, and zinc bioavailability by estimating the uptake, retention, and transport by Caco-2 cells.METHODS:Both dephytinized (by adding an exogenous phytase) and non-dephytinized infant cereals were digested using an in vitro digestion protocol adapted to the gastrointestinal conditions of infants younger than 6 mo. Mineral cell retention, transport, and uptake from infant cereals were measured using the soluble fraction of the simulated digestion and the Caco-2 cells. RESULTS: Dephytinization of infant cereals significantly increased (P<0.05) the cell uptake efficiency (from 0.66%-6.05% to 3.93%-13%), retention (from 6.04%-16.68% to 14.75%-20.14%) and transport efficiency (from 0.14%-2.21% to 1.47%-6.02%), of iron, and the uptake efficiency (from 5.0%-35.4% to 7.3%-41.6%) and retention (from 4.05%-20.53% to 14.45%-61.3%) of zinc, whereas calcium only cell uptake showed a significant increase (P<0.05) after removing phytate from most of the samples analyzed. A positive relationship (P<0.05) between mineral solubility and the cell uptake and transport efficiencies was observed.CONCLUSION: Removing phytate from infant cereals had a beneficial effect on iron and zinc bioavailability when infant cereals were reconstituted with water. Since in developing countries cereal-based complementary foods for infants are usually consumed mixed with water, exogenous phytase additions could improve the nutritional value of this weaning food.

The dynamics of the piglet gut microbiome during the weaning transition in association with health and nutrition

Background: Understanding the composition of the microbial community and its functional capacity during weaning is important for pig production as bacteria play important roles in the pig's health and growth performance. However,limited information is available regarding the composition and function of the gut microbiome of piglets in early-life.Therefore, we performed 16 S rRNA gene and whole metagenome shotgun sequencing of DNA from fecal samples from healthy piglets during weaning to measure microbiome shifts, and to identify the potential contribution of the early-life microbiota in shaping piglet health with a focus on microbial stress responses, carbohydrate and amino acid metabolism.Results: The analysis of 16 S rR NA genes and whole metagenome shotgun sequencing revealed significant compositional and functional differences between the fecal microbiome in nursing and weaned piglets. The fecal microbiome of the nursing piglets showed higher relative abundance of bacteria in the genus Bacteroides with abundant gene families related to the utilization of lactose and galactose. Prevotel a and Lactobacil us were enriched in weaned piglets with an enrichment for the gene families associated with carbohydrate and amino acid metabolism. In addition, an analysis of the functional capacity of the fecal microbiome showed higher abundances of genes associated with heat shock and oxidative stress in the metagenome of weaned piglets compared to nursing piglets.Conclusions: Overal, our data show that microbial shifts and changes in functional capacities of the piglet fecal microbiome resulted in potential reductions in the effects of stress, including dietary changes that occur during weaning.These results provide us with new insights into the piglet gut microbiome that contributes to the growth of the animal.

Supplemental effect of varying L-cysteine concentrations on the quality of cryopreserved boar semen

Cryopreservation 与反应的氧种类的生产被联系，它导致精子膜并且因而的类脂化合物 peroxidation 在精子活动性和减少的富饶潜力的减小。这研究的目的是决定为雄野猪精液的 cryopreservation 需要的 L 半胱氨酸的最佳的集中。12 只雄野猪为这研究提供了证明活动性和形态学的精液。精液被划分成乳糖鸡蛋蛋黄( LEY )扩展程序在使用了到的四部分重新推迟 centrifuged 精子小团与L半胱氨酸的各种各样的集中被补充到达 0 mmol L ? 1 (组我，控制)， 5 mmol L ? 1 (组 II )， 10 mmol L ? 1 (组 III )并且 15 mmol L ? 1 (组 IV )。精液暂停在稻草(0.5 mL ) 被装载并且在一个控制率冰箱放了。在 cryopreservation 以后，冻结精液样品为进步活动性，生存能力使用 SYBR-14/EthD-1 用染色的 FITC-PNA/EthD-1 染色和 acrosome 正直被融化并且调查。有一显著地更高(P < 0.01 ) 在与控制相比的二个 L-cysteine-supplemented 组(组 II 和组 III ) 的进步活动性，生存能力和 acrosomal 正直的百分比。有 L 半胱氨酸的 biphasic 效果，与在组 III 的进步活动性，生存能力和 acrosomal 正直的最高的百分比。在结论， 5 或 10 mmol L ? 1 是为改进冻结的鈥搕h awed 雄野猪精液的质量要加到 LEY 扩展程序的 L 半胱氨酸的最佳集中。

Detection and sequencing of plasmid encoded tetracycline resistance determinants(tetA and tetB) from food-borne Bacillus cereus isolates

Objective:To investigate the detection and sequencing of plasmid encoded tetracycline resistance genes(tet A and tet B) from food-borne and standard strains of Bacillus cereus(B. cereus).Methods:PCR was carried out to detect the tetracycline resistance genes(tet A and tet B) in food-borne B.cereus strains and the amplified products were sequenced.Results:The phenotypic resistance against tetracycline was observed in 39 of the 118 food-borne isolates and two reference strains(MTCC 430 and MTCC 1307) of B.cereus.Among the phenotypically resistant isolates,tet A was detected in 36 food-borne isolates and two reference strains(MTCC 430 and MTCC 1307).whereas,tel B was detected in 12 food-bome isolates and MTCC 1307 strain. Conclusions:A close association was therefore found between phenotypic resistance against tetracycline and presence of tetracycline resistance genes.The tet A and tet B gene fragments were amplified,purified and sequenced.The gene sequences of the isolates studied herein were found similar to tetA and tetB gene sequences of other bacteria available in NCBI.The occurrence of tetA and tetB genes in B.cereus indicate the horizontal transfer of antibiotic resistance determinants from other bacteria into B.cereus.The transfer of these resistant determinants to other potentially pathogenic bacteria may be a matter of great concern.

Development of polyurethane foam dressing containing silver and asiaticoside for healing of dermal wound

Polyurethane foam dressings for dermal wounds were formulated with natural polyols in order to improve the foam characteristics and the release of 2 active agents,silver and asiaticoside(AS)as an antimicrobial agent and an herbal wound healing agent,respectively.The foam was instantly formed by interaction of polyols and diisocyanate.Hydroxypropyl methylcellulose,chitosan and sodium alginate were individually mixed with themain polyols,polypropylene glycol,in the formulation while the active componentswere impregnated into the obtained foam dressing sheets.Although the type and amount of the natural polyols slightly affected the pore size,water sorption-desorption profile and compression strength of the obtained foam sheets,a prominent effect was found in the release of both active components.Among natural polyols formulations,foam sheets with alginate showed the highest silver and AS release.Non-cytotoxicity of these foam sheets to human fibroblast cells was confirmed.Antimicrobial testing on four bacteria strains showed that 1mg/cm^2 silver in formulations with 6%of natural polyols and without natural polyols had sufficient content of the silver release with comparable inhibition zone and significantly larger zone than other formulations.In pig study,the foam dressing with 6%alginate,1mg/cm^2 silver and 5%AS could improve wound healing in both the percentage of the wound closure and histological parameters of the dermal wound without any dermatologic reactions.In conclusion,this innovative foam dressing had potential to be a good candidate for wound treatment.

Seasonal abundance and potential of Japanese encephalitis virus infection in mosquitoes at the nesting colony of ardeid birds,Thailand

Objective:To investigate the abundance and seasonal dynamics of mosquitoes,and to detect Japanese encephalitis virus(JEV)in these mosquitoes at the nesting colony of ardeid birds.Methods:Mosquitoes were collected bimonthly from July 2009 to May 2010 by Centers for Disease Control.Light traps and dry ice,as a source of CO_2,were employed to attract mosquitoes.Mosquitoes were first identified,pooled into groups of upto 50 mosquitoes by species,and tested for JEV infection by viral isolation and reverse transcriptase polymerase chain reaction.Results:A total of 20370 mosquitoes comprising 14 species in five genera were collected.The five most abundant mosquito species collected were Culex tritaeniorhynchus(95.46%),Culex vishnui(2.68%),Culex gelidus(0.72%),Anopheles peditaeniatus(0.58%)and Culex quinquefasciatus(0.22%).Mosquito peak densities were observed in July.All of 416 mosquito pools were negative for JEV.Conclusions:This study provides new information about mosquito species and status of JEV infection in mosquitoes in Thailand.Further study should be done to continue a close survey for the presence of this virus in the ardeid birds.

Physicochemical properties and in vitro antioxidant activities of pyroligneous acid prepared from brushwood biomass waste of Mangosteen, Durian, Rambutan, and Langsat

Physicochemical characteristics and in vitro antioxidant activities of four pyroligneous acids carbonized from the wastes of wood species including Mangosteen (Garcinia mangostana Linn.), Durian (Durio zibethinus L.), Rambutan (Nephelium lappaceum L.), and Langsat (Lansium domesticum Serr.) were assessed. Appearing as transparent liquors with pH 3.9–4.2, the pyroligneous acid samples under test possessed acetic acid (23.22–25.46%) as the dominant component. The total soluble tar, total acid, and water content were 0.15 - 0.28 wt%, 99–192 mg KOH/g and 84.5–93.5 wt%, respectively. Phenolic compounds namely: 2,6-dimethoxyphenol (6.88–9.69%),phenol (2.97–5.88%), 4-methylsyringol (3.10–3.56%), guaiacol (2.36–3.55%), and 2-methoxy-4-methylphenol (1.08–1.28%) were found. All had in vitro antioxidant activities especially mangosteen pyroligneous acid, which showed activity roughly similar to BHT (P>0.05) against anti-lipid peroxidation. Nitric oxide scavenging capacities of all pyroligneous acids were significantly higher than BHT (P<0.05). Our results suggest that pyroligneous acids from the four types of branch waste could be used as sources of beneficial natural antioxidants, possibly as food or feed additives to protect against lipid peroxidation, and potentially also in veterinary medicine in anti-inflammatory products.

Comment on: Cloning and characterization of porcine aquaporin 1 water channel expressed extensively in the gastrointestinal system

TO THE EDITOR Sir,I read with great interest the recently pubiished article in rhe World Journal of Gastroenterology by JIn and co-workers on the cloning and characterization of Porcine aquaporin 1 water channel from the pig liver and studies on its expression in

Detection of Babesia bovis in blood samples and its effect on the hematological and serum biochemical profile in large ruminants from Southern Punjab

Objective:To determine the presence of Babesia bovis(B.bovis)in large ruminants in southern Punjab and its effect on hematological and serum biochemical profile of host animals.Methods:Blood samples were collected from 144 large ruminants,including 105 cattle and 39 buffaloes,from six flistricts in southern Punjab including Multan,Layyali,Muzaffar Garh,Bhakar.Bahawalnagar and Vehari.Data on the characteristics of animals and herds were collected through questionnaires.Different blood(hemoglobin,glucose)and serum(ALT.AST,I.DH,cholesterol)parameters of calves and cattle were measured and compared between parasite positive and negative samples to demonstrate the effect of B.bovis on the blood and serological profile of infected animals.Results:27 out of 144 animals,from 5 out of 6 sampling districts,produced the541-bp fragment specific for B.bovis.Age of animals(P=0.02),presence of ticks on animals(P=0.04)and presence of ticks on dogs associated with herds(P=0.5)were among the major risk factors involved in the spread of bovine babesiosis in the study area.ALT concentrations were the only serum biochemical values that significantly varied between parasite positive and negative cattle.Conclusions:This study has reported for the first time the presence of B.bavis in large ruminant and the results can lead to the prevention of babesiosis in the region to increase the livestock output.

Detection and characterization of Chlamydophila psittaci in asymptomatic feral pigeons(Columba livia domestica) in central Thailand

Objective:To detect and characterize Chlamydophila psittaci(C.psittaci) in asymptomatic feral pigeons in central Thailand.Methods:A total 814 swabs from the trachea and cloacae of 407non-clinical feral pigeons in central Thailand were collected and tested for the presence of C.psittaci.Results:A 10.8%of feral pigeons in the sample group were positive as determined by nested PCR primer specific to C.psittaci.The outer membrane protein A(orupA) gene of positive samples exhibited amino acid identity of C.psittaci ranging from 71 to 100%and were grouped in genotype B.Exceptionally,BF1676-56 isolate was closely related to Chlamydia avium with99%identification of the I6 S ribosomal(r) RNA gene.Conclusions:This is the first report on C.psittaci isolated from asymptomatic feral pigeons in Thailand,which provides knowledge for the disease status in pigeon populations in Thailand.

Production of spike and nucleocapsid recombinant proteins of porcine epidemic diarrhea virus for antibody detection by ELISA

Porcine epidemic diarrhea (PED), a devastating enteric disease in pigs, is caused by PEDvirus (PEDV)(1)Reduced severity of clinical diseases was reported to associate with neutralizing antibody titers in colostrum. However, viral neutralization assay(VN) is laborious and not suitable for routine diagnosis. Spike protein plays an important role in stimulating neutralizing antibody that might be suitable for PEDV diagnosis.

Terminalia arjuna:A novel natural preservative for improved lipid oxidative stability and storage quality of muscle foods

The study was conducted to explore the possibility of utilization of Terminalia arjuna as a novel natural preservative in meat products by using chevon sausages as a model system.Chevon sausages were prepared by incorporating different levels of T.arjuna viz.T1(0.25%),T2(0.50%)and T3(0.75%)and were assessed for various lipid oxidative stability and storage quality parameters under refrigerated(4±1◦C)conditions.T.arjuna showed a significant(p<0.05)effect on the lipid oxidative stability as the treated products exhibited significantly(p<0.05)lower TBARS(mg malonaldehyde/kg)values in comparison to control.A significant(p<0.05)effect was also observed on the microbial stability as T.arjuna incorporated products showed significantly(p<0.05)lower values for total plate count(log cfu/g),psychrophilic count(log cfu/g),yeast and mould count(log cfu/g)and FFA(%oleic acid)values.Significantly(p<0.05)higher scores were observed for various sensory parameters of the products incorporated with T.arjuna during refrigerated storage.T.arjuna successfully improved the lipid oxidative stability and storage quality of the model meat product and may be commercially exploited as a novel preservative in muscle foods.

Microsatellite-Based Genetic Differentiation and Phylogeny of Sheep Breeds in Mongolia Sheep Group of China

In the present study we studied the genetic structure of five Chinese sheep populations of Hu sheep, Tong sheep, Small-tailed Han sheep, Tan sheep, and Wadi (WD) sheep using 15 microsatellite loci. The results showed that the FIT, FST, and FIS statistics computed for the complete dataset had the following values, 0.523±0.140, 0.363±0.131 and 0.263±0.092, respectively. All loci were significantly contributed to the genetic differentiation among population (P<0.001). There is no relationship between the scatter of pairwise FST geographical distance points as geographical distance increases between the five populations. Membership probabilities and genetic structure of sheep populations were estimated when K=2, the populations were classified into Hu, Tong, Han and WD, and Tan sheep group. However, when K=3, the populations were classified into Hu and Tong, Han and WD, and Tan sheep group. The findings supported the previous literatures that these populations are originated on different time stages from the primogenitor population and communicated genetically with each other by natural and artificial selection in different ecological environment.

Embryoid body formation from embryonic and induced pluripotent stem cells:Benefits of bioreactors

Embryonic stem(ES)cells have the ability to differ-entiate into all germ layers,holding great promise not only for a model of early embryonic development but also for a robust cell source for cell-replacement therapies and for drug screening.Embryoid body (EB)formation from ES cells is a common method for producing different cell lineages for further applications. However,conventional techniques such as hanging drop or static suspension culture are either inherently incapable of large scale production or exhibit limited control over cell aggregation during EB formation and subsequent EB aggregation.For standardized mass EB production,a well defined scale-up platform is necessary.Recently,novel scenario methods of EB formation in hydrodynamic conditions created by bioreactor culture systems using stirred suspension systems(spinner flasks),rotating cell culture system and rotary orbital culture have allowed large-scale EB formation.Their use allows for continuous monitoring and control of the physical and chemical environment which is difficult to achieve by traditional methods.This review summarizes the current state of production of EBs derived from pluripotent cells in various culture systems.Furthermore,an overview of high quality EB formation strategies coupled with systems for in vitro differentiation into various cell types to be applied in cell replacement therapy is provided in this review. Recently,new insights in induced pluripotent stem(iPS) cell technology showed that differentiation and lineage commitment are not irreversible processes and this has opened new avenues in stem cell research.These cells are equivalent to ES cells in terms of both self-renewal and differentiation capacity.Hence,culture systems for expansion and differentiation of iPS cells can also apply methodologies developed with ES cells,although direct evidence of their use for iPS cells is still limited.

Ethanol consumption as inductor of pancreatitis

Alcohol abuse is a major cause of pancreatitis, a condition that can manifest as both acute necroinflammation and chronic damage (acinar atrophy and f ibrosis). Pancreatic acinar cells can metabolize ethanol via the oxidative pathway, which generates acetaldehyde and involves the enzymes alcohol dehydrogenase and possibly cytochrome P4502E1. Additionally, ethanol can be metabolized via a nonoxidative pathway involving fatty acid ethyl ester synthases. Metabolism of ethanol by acinar and other pancreatic cells and the consequent generation of toxic metabolites, are postulated to play an important role in the development of alcohol-related acute and chronic pancreatic injury. This current work will review some recent advances in the knowledge about ethanol actions on the exocrine pancreas and its relationship to inflammatory disease and cancer.

Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease

Enzyme-linked immunosorbent assay (ELISA) is often used to test bovine leukemia virus (BLV) infection. However, commercially available kits test in South America detect only antibodies against the gp51 protein. With the aim to improve the sensitivity of the test, we developed here a two-step indirect dual ELISA test that included both proteins p24 and gp51, expressed and produced in E. coli and baculovirus expression system respectively. Two hundred ten BLV sera, stated as double positive or double negative by the combination of commercial agar gel immunodiffusion (AGID) assay and a gp51-ELISA test, were tested with our in house dual rp24/rgp51 ELISA. Firstly, we checked the purified, optimized and standardized proteins as antigen by the checkerboard technique, and set up our in house ELISA test. The concordance correlation coefficient (CCC) and coefficient of variation (CV) intraplate repeatability levels were within the values established by the international standards. The statistical analysis demonstrated the value of sera correctly ranked highest (93.48%), and for 0.3 cutoff, the sensitivity was 95.65% and the specificity 91.30%. In conclusion, the rp24/rgp51 ELISA developed and standardized here demonstrated to have good analytical characteristics to be considered for screening of BLV.

Manipulation of bicarbonate concentration in sperm capacitation media improves in vitro fertilisation output in porcine species

Background: The in vivo concentration of bicarbonate(HCO3-), one of the essential sperm capacitating effectors,varies greatly in the different environments sperm go through from cauda epididymis to the fertilisation site. On the contrary, porcine in vitro sperm capacitation and fertilisation media usually contains a standard concentration of25 mmol/L, and one of the main problems presented is the unacceptable high incidence of polyspermy. This work hypothesised that by modifying the HCO3-concentration of the medium, the output of in vitro sperm capacitation and fertilisation could be increased.Results: Once exposed to the capacitation medium, the intracellular pH(pHi) of spermatozoa increased immediately even at low concentrations of HCO3-, but only extracellular concentrations of and above 15 mmol/L increased the substrates protein kinase A phosphorylation(pPKAs). Although with a significant delay, 15 mmol/L of HCO3-stimulated sperm linear motility and increased other late events in capacitation such as tyrosine phosphorylation(Tyr-P) to levels similar to those obtained with 25 mmol/L. This information allowed the establishment of a new in vitro fertilisation(IVF)system based on the optimization of HCO3-concentration to 15 mmol/L, which led to a 25.3% increment of the viable zygotes(8.6% in the standard system vs. 33.9%).Conclusions: Optimising HCO3-concentrations allows for establishing an IVF method that significantly reduced porcine polyspermy and increased the production of viable zygotes. A concentration of 15 mmol/L of HCO3-in the medium is sufficient to trigger the in vitro sperm capacitation and increase the fertilisation efficiency in porcine.