Supplementation of L-ascorbic acid improves the in vitro development of buffalo(Bubalus bubalis)embryos and alters the expression of apoptosis-related genes

Objective:To study the effect of L-ascorbic acid supplementation on the in vitro development of buffalo embryos and evaluate the relative mRNA abundance of some pro-apoptotic,anti-apoptotic,and embryonic development-related genes.Methods:In experiment 1,we evaluated the effect of the addition of 0(control),50,and 100μM L-ascorbic acid to the in vitro maturation medium on the developmental competence in terms of blastocyst rate and relative mRNA abundance of some pro-apoptotic(BAX,BID),anti-apoptotic(BCL-XL,MCL1),and embryonic development(GDF9,BMP15)related genes.Based on the results,we chose 50μM as the suitable dose of L-ascorbic acid for the subsequent experiments.We further evaluated the blastocyst rates following the addition of 50μM L-ascorbic acid to the in vitro culture medium(experiment 2),and in vitro maturation and in vitro culture media(experiment 3).In all three experiments,the maturation and culture media devoid of L-ascorbic acid served as the control group.Results:The blastocyst rate after adding 50μM L-ascorbic acid to the in vitro maturation medium was significantly higher than the control group(P<0.05),whereas 100μM L-ascorbic acid exhibited a negative effect on the blastocyst rate.The blastocyst rates for embryos cultured in 50μM L-ascorbic acid in the in vitro culture medium alone and both in vitro maturation and in vitro culture media were significantly higher than their corresponding control groups(P<0.05).The relative mRNA abundance of BAX significantly decreased in blastocysts produced after the addition of 50μM L-ascorbic acid as compared with the control group(P<0.05),whereas,for MCL1,it significantly decreased in blastocysts produced after the addition of 100μM L-ascorbic acid(P<0.05).Conclusions:The supplementation of 50μM L-ascorbic acid to in vitro maturation and in vitro culture media supports in vitro embryonic development in buffaloes by improving developmental competence and altering the expression of apoptosis-related genes.

Identification of stable internal control genes for accurate normalization of real-time quantitative PCR data in testicular tissue from two breeds of cattle

Objective:To assess the stability of 10 candidate internal control genes(ICGs),namely GAPDH,ACTB,RPL23,RPS15A,ATPSF1,GLUT5,HMBS,ATP2B4,PPIA,and BRP to normalize the transcriptional data from testes samples of Zebu and crossbred bulls.Methods:Total RNA was isolated from testicular tissue of Zebu and crossbred bulls(n=6 each)between 2-8 years of age.cDNA was synthesized,and the quantitative real-time polymerase chain reaction(PCR)was performed.The cycle threshold values were used for the analysis of the stability of ICGs.Four different statistical algorithms:geNorm,Normfinder,BestKeeper,and RefFinder,were used to assess the stability of these genes.Results:ATPSF1,HMBS,PPIA,and RPS15A were the most reliable and stable ICGs for Zebu testes,and ATPSF1,RPL23,and PPIA for crossbred testes.Conclusions:A panel of stable ICGs(ATPSF1,HMBS,PPIA,RPS15A for Zebu and ATPSF1,RPL23,and PPIA for crossbred)for normalization of gene expression data in testes samples can be helpful for researchers to conduct functional genomics studies at the testicular level in cattle bulls.

Role of preputial washing in reducing microbial load and improving bovine semen quality

Quality semen production remains the main focus and objective of semen processing laboratories throughout the world. Bacterial and other microbial contaminants affect the semen quality and hence the fertility, and also lead to reproductive disorders as well as lower conception rates and increased embryonic mortality, abortion and other complications in females. Microbial contamination affects the semen adversely, by exerting direct spermicidal effect, formation of reactive oxygen species, toxin production, adherence with spermatozoa, deriving nutrients and oxygen from the medium and thus competing with spermatozoa for the factors of growth and normal functioning. Despite hygienic measures, several ubiquitous and opportunistic microbes find their ways into semen during collection, processing, and storage of semen, and survive even during freezing. Stringent sanitary precautions are therefore required at every step of collecting semen and its processing. Preputial cavity is considered as main source of semen contaminating microorganisms. Flushing the preputial cavity with normal saline or any suitable liquid combination with antimicrobial activity, prior to semen collection reduces the microbial load and thereby improves the semen quality.

Sperm dosage and site of insemination in relation to fertility in bovines

Low sperm numbers in artificial insemination (AI)-doses are being used widely to make the best use of high genetic value bulls as well as sex-sorted semen. Sperm concentration needed for AI to obtain reasonable fertility, taking genetic value of bull and numerous others components into consideration is one of the essential constituents for successful AI breeding program. However, low sperm concentrations in AI-doses lead to reducing post-thaw viability. The reduction in viability of low sperm doses may be affected by fresh semen volume, sperm number and seminal plasma level at final dilution. Reduction in quality and fertility of low sperm doses is one of the limitations for their use in successful AI programme. Sperm number per AI required to achieve optimum fertility is one of the main crucial things to AI industry, and numerous efforts have been made in this regard. Due to great variability among bulls, sperm number per AI could be a limiting factor in achieving acceptable fertility values. Fertility of low sperm doses may vary among bulls, and non-return rates (NRRs) with low sperm doses may be determined by fertility level of bull. On the basis of individual bulls, sperm numbers in AI doses needed to be adjusted to reduce the variations in NRRs among bulls. Utilizing high fertile bulls for low sperm doses with acceptable non-return rates (NRRs) may be a way to cover a large number of bovines under AI in countries like India. Deposition site within the uterine horn may alter non return rates following inseminations with low sperm doses. Following deep-uterine inseminations, acceptable pregnancies may be achieved with low sperm doses and even if ovulation side is unknown.

Laser irradiation effects and its possible mechanisms of action on spermatozoa functions in domestic animals

This article presents a review pertains the laser irradiation effects and its possible mechanisms of action on spermatozoa functions in domestic animals. To improve artificial insemination, laser is sensitive and cost effective technique, when compared to other conventional methods. Laser may have both positive and negative effects on spermatozoa functions. Since the effects of light are mediated by reactive oxygen species, and the levels of these reactive oxygen species following irradiating spermatozoa with laser may be responsible for determining the effects of laser on sperm. Dose of laser may be regarded as of great significance and this dosage of laser may be responsible for determining its effects on spermatozoa. Optimum dosage of laser for improving seminal attributes may vary among various species and this need to be standardized in each of them. The beneficial effects include improving sperm livability, acrosomal integrity, hypo-osmotic swelling response, mitochondrial function and computer-aided sperm analysis parameters. The increase in cytochrome c oxidase activity, ATP levels and mitochondrial membrane potential, in laser irradiated cells may be responsible for enhanced sperm quality parameters. Improving fertility with laser irradiated spermatozoa has been reported in few species like boar and need to be elaborated in other species. In conclusion laser may be regarded as an easy, cheap and time saving technology for improving artificial insemination;in addition, laser may have various potential applications in the field of reproductive biotechnology as well as in livestock farms and veterinary polyclinics.

Prospects of diagnostic and prognostic biomarkers of pyometra in canine

Pyometra is one of the most common uterine pathologies of intact bitch at middle to advanced age. In the early stages, the disease shows subtle changes, making diagnosis a challenge. In contrast, at later stages, it manifests as potentially life-threatening systemic inflammatory response syndrome. Ultrasonographic examination of the uterus aids in the diagnosis, although it has limitation in ascertaining the clinical severity of pyometra. Moreover, differentiation of cystic endometrial hyperplasia from pyometra could not be discerned with greater accuracy. Therefore, false negative diagnosis of pyometra patients leads to development of systemic inflammatory response, which delays administration of therapies and results in deaths during early course of treatment. Further, indiscriminate use of broad-spectrum antimicrobials at higher dose in false positive cases considerably contributes to the rising pool of drug resistant pathogens, thereby increasing the risk of case fatality due to sepsis in a long-term. Monitoring the circulating pro-inflammatory cytokines, acute phase proteins, endotoxin, growth factors and inflammatory mediators is the current trend in pyometra research, especially for developing diagnostic and prognostic biomarkers. The present review deals with the prospects of developing diagnostic and prognostic biomarkers in the canine pyometra.

Recent developments in dairy kefir-derived lactic acid bacteria and their health benefits

Kefir is one of the most consumed traditional fermented dairy products and it has a complex probiotic and nutritional composition.Kefir grain contains casein and other milk solids together with the yeasts and lactobacilli that cause the characteristic kefir fermentation and serve as a starter to induce this fermentation when introduced into fresh milk.Both kefir and kefir grains have a rich microbiota and their composition is affected by many parameters like kefir origin,production method(artisanal or industrial),kefir grain:milk ratio,type of milk,fermentation conditions,inoculum source,equipment used in production and storage conditions.Lactic acid bacteria(LAB)are a significant part of the kefir’s microbial composition and the health-promoting effects.Kefir-derived LAB have beneficial effects on colorectal cancer,cardiovascular diseases,type 2 diabetes mellitus,obesity,kidney diseases,modulation of the immune system as well as intestinal microbiota through different biological mechanisms.In addition,bioactive peptides and metabolic products of kefir have shown promising results as health beneficial components along with certain antiviral effects including COVID-19.Therefore,this article is up-to-date information about the production,processing as well as health benefits of dairy kefir-derived LAB with future prospect of work.

Nano zinc, an alternative to conventional zinc as animal feed supplement:A review

The uniqueness of Zn is that, it is the second most abundant trace element in the animal body but can't be stored in the body, thus regular dietary intake is required. Zinc oxide(ZnO) nanoparticles(NP) particles are being extensively used in paints, skin lotions pigments, food, electronics appliances, biological and pharmaceutical applications and many more. Zinc oxide nanoparticles are the specially prepared mineral salt having particle size of 1 to 100 nm. It promotes growth can act as antibacterial agent,modulates the immunity and reproduction of the animals. Both in lower and higher doses of specifications it has exhibited a variety of effects on animal performances. Apart from being highly bioavailable, reports have already pointed out the growth promoting, antibacterial, immuno-modulatory and many more effects of nano zinc(nZn). These can be used at lower doses and can provide better result than the conventional Zn sources and indirectly prevents environmental contamination also. The toxicological studies provide mixed results in animal models. Studies been undertaken in diversified animal species and encouraging effects have been reported with nZn supplementation. However, there is a need to optimize the dose and duration of ZnO NP supplementation for human and livestock,depending on its biological effects. Actual bioavailability of ZnO NP in livestock is still to be worked out.In this review we have attempted to summarize, conclude the beneficial effects of nZnO and its possible usage as mineral supplement to different categories of human and livestock.

Growth performance, and enteric and manure greenhouse gas emissions from Murrah calves fed diets with different forage to concentrate ratios

The present study investigated the effects of different dietary forage to concentrate ratios on animal performance, and enteric and manure greenhouse gas emissions in growing calves.Fifteen male Murrah calves(153.5 ± 18.17 kg; 6 to 12 months) were randomly assigned to 3 dietary treatments and fed corn fodder, wheat straw and concentrate in 3 different proportions: 20:60:20(C20); 20:40:40(C40) and 10:30:60(C60), for a period of 120 days.Increasing dietary concentrate proportion had no significant(P > 0.05) effect on dry matter intake(DMI) but increased crude protein(CP) and total digestible nutrient intake(P < 0.05).Average daily gain and feed conversion efficiency were significantly higher(P < 0.05)for C60 compared with C20 and for C40, these did not differ with C20 and C60(P > 0.05).The apparent digestibility of dry matter, organic matter and CP were higher(P < 0.05), but acid detergent fiber digestibility was lower(P < 0.05) for C60 compared with C20 whereas, ether extract and neutral detergent fiber digestibilities were not affected(P > 0.05).Daily methane(CH_4) emission(g/d), CH_4 energy loss(MJ/d) and CH_4 yield(CH_4 g/kg organic matter intake [OMI], CH_4 g/kg digestible OMI, and CH_4% of metabolizable energy intake) were significantly higher for C20 compared with C60(P < 0.05).Methane yield as g/kg DMI although lower for C60 compared with C20 but the difference was not significant(P > 0.05).Manure CH_4(g/kg DMI) and nitrous oxide(N_2O mg/kg nitrogen) emissions were not affected(P > 0.05), but N_2O emission on mg/kg DM basis was significantly higher(P < 0.05) from the manure of calves fed C60 than that for C20.Thus, increasing dietary concentrate proportion improved animal performance, and reduced enteric CH_4 emission(g/day) without any significant effect on manure N_2O(mg/kg nitrogen) and CH_4 emissions.

A clinically relevant combination treatment with doxorubicin and cyclophosphamide does not induce hepatotoxicity in C57BL/6J mice

Background and aims:Chronic exposure to chemotherapeutics can lead to severe adverse events including hepatotoxicity.A combination chemotherapy regimen of doxorubicin(DOX)and cyclophos-phamide(CPS)is employed in treatment of several cancers such as leukemia,lymphoma,and breast cancer.It is not well understood whether a combination therapy with DOX and CPS can induce hepa-totoxicity.We therefore sought to determine whether co-administration of DOX and CPS at their clini-cally relevant doses and frequency results in hepatotoxicity.Methods:Male C57BL/6J mice received one intraperitoneal injection of saline or DOX-2 mg/kg and CPS-50 mg/kg once a week for 4 weeks.After the treatment period,liver histology and various serum bio-markers of hepatotoxicity were assessed.Results:Co-treatment with DOX and CPS did not alter the serum levels of alanine aminotransferase(ALT),alkaline phosphatase(ALP),bilirubin,albumin,globulin,or total protein.Similarly,co-administration of DOX and CPS did not result in a noticeable change in liver histology.However,it was notable that the concomitant treatment with DOX and CPS resulted in a significant increase in serum levels of aspartate aminotransferase(AST).Elevated serum AST levels were also associated with increased serum creatinine kinase(CK)levels,suggesting that the elevated serum AST levels are likely due to muscle injury following the co-administration of DOX and CPS.Conclusions:Taken together,our results,for the first time,suggest that co-administration of DOX and CPS,at their clinically relevant doses and frequency does not induce a significant hepatotoxicity in the mice.

Stability assessment of emulsion of carotenoids extracted from carrot bio-waste in flaxseed oil and its application in food model system

The research was conducted on the stability assessment of emulsion of carotenoids extracted from carrot-pomace in omega-3 rich flaxseed oil,followed by converting this emulsion into freeze-dried powder,and its application as natural functional colorant in the preparation of flavored milk.The study indicated that emulsion was stable over a range of pH (5–9),temperature (forewarming to HTST pasteurization) and ionic strength (0.1–0.75 M NaCl) except at pH 4,sterilization temperature and 1.0 M NaCl based on zeta potential and particle size of emulsion analysed at these conditions.Ten percent (out of 8,10 and 12%) of freeze-dried emulsion was selected for addition as a natural functional ingredient in flavored milk on the basis of organoleptic evaluation.The fat,protein,total solids,total carotenoid content and antioxidant value (in terms of ABTS and reducing power assay) of developed product were significantly higher (p < 0.05) than control.The whiteness index of control was significantly higher (p < 0.05) than the developed product,while reverse was true for yellowness index.However,both the products showed total color difference of 34.65.The research concludes that carotenoid-rich emulsion can be efficiently freeze-dried and used as a functional natural colorant in flavored milk for providing it better bio-functional attributes over control.

Nitrogen fractionation of certain conventional-and lesser-known by-products for ruminants

Dietary proteins for ruminants are fractionated according to solubility, degradability and digestibility. In the present experiment, 11 vegetable protein meals and cakes used in ruminant nutrition were included with a main focus on determining various nitrogen(N) fractions in vitro. Total N(N×6.25) content varied from 22.98%(mahua cake) to 65.16%(maize gluten meal), respectively. Guar meal korma contained the lowest and rice gluten meal had the highest acid detergent insoluble nitrogen(ADIN; N×6.25). Boratephosphate insoluble N(BIN, N×6.25) and Streptomyces griseus protease insoluble N(PIN; N×6.25) were higher(P<0.01) in maize gluten meal than in other feeds, whereas groundnut cake and sunflower cake had lower(P < 0.01) BIN, and PIN, respectively. Available N, calculated with the assumption that ADIN is indigestible, was maximum in guar meal korma and minimum in rice gluten meal. Furthermore, rapid and slowly degradable N(N × 6.25) was found to be higher(P < 0.01) in groundnut cake and coconut cake, respectively. Intestinal digestion of rumen undegradable protein, expressed as percent of PIN, was maximum in guar meal korma and minimum in rice gluten meal. It was concluded that vegetable protein meals differed considerably in N fractions, and therefore, a selective inclusion of particular ingredient is needed to achieve desired level of N fractions to aid precision N rationing for an improved production performance of ruminants.

Social network structures among the livestock farmers vis a vis calcium supplement technology

This study aims to delineate and analyze the configuration of social networks of farmers with respect to the acquisition of information on vital livestock technology.Three stage sampling was carried out by interviewing 320 technology-adopter farmers from four districts of Kerala State in India.For mapping the network,social network analysis(SNA)was used,which revealed the important sources as well as patterns of information access by farmers.Results established the predominance of a formal communication source(veterinary doctor)in the study locales followed by small-sized peer groups of livestock farmers for crucial information support on technology use.The trend is observed irrespective of their gender in various study areas.Significantly the study thus,underscored the role of homogenous peer groups of farmers in facilitating meaningful interactions as well as information sharing on the technology.Given the low level of adoption of most livestock technologies along with the weak livestock extension machinery in the country,these findings could be used by extension agencies to strategize future technological interventions.

Production of whey protein hydrolyzates and its incorporation into milk

Whey proteins provide an excellent source of low-molecular-weight bioactive peptides with important functional properties and bioactivities like antihypertensive,opioid,and antimicrobial effects.Presence of peptide molecules with lower molecular weight has a great role in food for health promotion.In this investigation,the release of low-molecular-weight peptides from whey protein concentrate was attempted by using enzymatic digestion.The hydrolyzate was then incorporated into milk to obtain enriched milk(EM)with low-molecular-weight peptides.Based on sensory analysis of EM,electrophoretic and RP-HPLC studies,hydrolyzates of 10%protein(degree of hydrolysis 5%;enzyme/substrate E/S,1:50)were finally incorporated into milk at 20%(v/v)to develop an acceptable product enriched with low-molecular-weight peptides.EM had higher protein content,viscosity and emulsifying properties than control milk with 3%fat.It is recommended that EM should not be sterilized as it results in coagulation,but can be safely pasteurized and spray dried without any undesirable effects.Maximum ACE-inhibition activity was obtained in hydrolyzate,followed by EM.This study is expected to boost the opportunity for the dairy industry to venture further into the nutraceutical dairy market.