The bovine kappa-casein (κ-CN) is a phospho-protein with 169 amino acids encoded by the CSN3 gene. The two most common gene variants in the HF breed are CSN3*A and CSN3*B while CSN3*E has been found with lower frequency. The aim of this study was to optimize a laboratory method for genotyping of these three alleles as well as to determine their genotype and allele frequencies in the HF cattle population in the Republic of North Macedonia. Genomic DNA was extracted from full blood from 250 cows. The target DNA sequence was amplified with newly designed pair of primers and the products were subjected to enzymatic restriction with HindIII and HaeIII endonucleases. Genotype determination was achieved in all animals. The primers successfully amplified a fragment of 458 bp and the digestion of this fragment with both endonucleases enabled differentiation of five different genotypes with the following observed frequencies: AA (0.39), AB (0.29), BB (0.16), AE (0.10), and BE (0.06). The estimated allele frequencies were: CSN3*A (0.584), CSN3*B (0.336) and CSN3*E (0.08). The observed genotype frequencies differed significantly (P<0.01) from those that would be expected under HW equilibrium, while the fixation index (F=0.17) indicated moderate heterozygosity deficiency. Nevertheless, the CSN3*B allele was present with relatively high frequency which should be used to positively select for its carriers, since increasing its frequency could help to improve the rheological properties of the milk intended for cheese production.
Chicken astroviruses (CAstV) are enteric viruses of poultry causing gastroenteritis, malabsorption, gout and white chick disease commonly known as runting-stunting syndrome (RSS). It can affect the wide range of poultry birds, especially chicken, turkey and duck worldwide. To our best knowledge there is no published report on presence of antibodies against CAstV in Bangladesh. Therefore, the study aimed to detect the presence of CAstV antibodies in broilers and sonali chickens (a cross-bread) in Bangladesh through a cross-sectional survey. A total of 454 blood samples from 66 flocks of broiler (n=343) and sonali chickens (n=111) of different ages were obtained during 2017 from four districts. The birds were healthy but were not vaccinated against CAstV. The samples were tested for specific antibodies against CAstV Group B by using commercially available ELISA kit. Overall, 16.74% (76/454) samples and 34.84% (23/66) flocks were positive for CAstV antibodies. The seroprevalence of CAstV was significantly (p=0.001) higher in sonali chickens (36.96%) than broiler (10.20%), while it was significantly higher (p=0.001) in birds of Bogura district (36.94%) than the other three districts. Regarding the age groups, seroprevalence was insignificantly (p=0.192) higher in sonali chicken before laying age (45%) than during laying age (27.45%). Regarding the seasons, CAstV infection was prevalent significantly (p=0.001) higher in winter season. Thus, the present study indicated the presence of CAstV in poultry in Bangladesh, so further studies are required to find out the magnitude of the problem in the country.
Diffuse cutaneous mastocytosis was diagnosed in a 6-year-old, indoor, neutered female domestic European shorthair cat. Marked pruritus located mainly on the head and neck was noticed in the cat and in this area the animal had developed alopecia, crusts, and plaques. Histologically, monomorphic mast cells were found in the superficial dermis and around the hair follicles. Mast cells were well differentiated, with central nuclei and granular cytoplasm, with metachromatic granules which stained positively with Toluidine blue stain. The animal was successfully treated with oclacitinib at a dose of 1 mg/kg, twice a day per os.
In recent years many researchers have described the reduced mycotoxin toxicity caused by probiotic bacteria. Since reduction under gastrointestinal conditions of the bioavailability of mycotoxins by probiotics is not fully investigated in birds, the aim of the study was to determine the effect of probiotic on T-2 mycotoxicosis induced apoptosis in broiler’s liver. For the study, twelve 1-days old broilers were divided equally into T-2 toxin (T2) and probiotic with T-2 (P+T2) groups. From the first experimental day, probiotic Enterococcus faecium DSM 7134 was administered in drinking water to P+T2 group. From the fourth day, T-2 toxin was given for three consecutive days to T2 toxin group. At 8th experimental day chicken were sacrificed, liver was fixed in buffered 10% formalin, embedded into paraffin, slices 5 μm in thickness were cut followed by immunohistochemical staining with polyclonal primary antibodies p21 and p53 (Abcam, UK) according to the manufacturers’ guidelines (IHC kit, Abcam, UK). Blood samples were taken by cardiac puncture to measure liver enzymes. Immunohistochemical staining revealed strong expression of p53 and p21 antibodies in hepatocytes nuclei as well as around blood vessels in T-2 toxin group’s chicken liver tissue. Staining by both antibodies was less intensive in P+T2 group. Enzyme analysis showed significantly increased (p<0.05) blood aspartate transaminase and alanine transaminase concentrations by 33.87% and 68.03% respectfully in T2 toxin group, while enzyme concentrations were decreased in P+T2 group. The obtained results showed reduced features of liver apoptosis in treatment with probiotic bacteria.
The present study aimed to evaluate whether the induction and the formation of an accessory corpus luteum (CL) after AI might increase the pregnancy per AI (P/AI) in heat stressed dairy cows. Starting at d 50±3 post-partum, 113 lactating Holstein cows from one commercial herd during summer were scored for body condition, blood sampled and examined by ultrasound. Those bearing a CL>25mm and progesterone (P4) level>2ng/mL were synchronized using a double PGF2α injection given 12 h apart and AI-ed at detected estrus. In total 18 cows, there were not any signs of estrus (n=10) nor a P4 level <2ng/mL at the time of enrolment (n=8) and therefore they were excluded from the study, leading to 95 cows finally enrolled.. At d5 post-AI, cows were randomly allocated into 2 groups: control group (CON, n=45) without any additional treatment, and treatment group (GnRH, n=50), treated with 0.008 mg Buserelin – a GnRH agonist. Blood sampling and ultrasound examination were done at d5, d14 and at d21 after AI, whereas the pregnancy diagnosis was done at d21 and d30 after AI. Average daily temperature and relative humidity values were used to calculate the temperature-humidity index (THI). The average THI during the experiment was 79.5±0.6. At d5, no differences were observed neither between the number of the CL nor between the P4 level in both groups. At d14 and d21, 82% of the GnRH-treated cows had more than one CL versus 0% of the CON cows. Both at d14 and 21, GnRH-treated cows had higher P4 levels compared to the CON cows (p<0.05). In addition, P/AI were higher in the GnRH group than in the CON group (65% vs. 48.3%, p<0.05), whereas late embryonic losses were higher in CON in comparison to GnRH cows (10.6 vs. 4%, respectively). The BCS at the moment of insemination did not affect P/AI (p>0.05). In conclusion, the induction of an accessory CL at d 5 after AI might increase P/AI in heat stressed dairy cows.
Since there is not enough data about milkability of the Jersey cows, the aim of this paper is to show basic milkability traits of this cattle breed depending on the parity and milk flow curve types. Cows had average daily production (DMY) of 22.23 kg, milk yield per milking (MYM) of 9.72 kg, and average and maximum milk flow about 1.66 and 2.49 kg/min, respectively. Statistical analysis showed that DMY and MYM of the cows in 4th parity was significantly (p<0.05) higher comparing to cows in 1st and 3th parity. The duration of entirely milking is similar regarding the paritiesand only the cows in 3th and 4th differ significantly (p<0.05). Regarding the effect of milk flow curve, the significant (p<0.05) difference between unspecified and rectangular milk flow curve has been found for the average milk flow (AMF) and descending phase of the milk flow curve (TD). Cows with unspecified milk flow curve, have significantly (p<0.05) lower AMF and longer TD compared to cows with rectangular curve. Cows with bimodal milk flow curve have significantly (p<0.05) longer duration of the ascending phase compering to: unspecified, descending and rectangular. The results of this study have shown that Jersey cows have lower production and milk flow compared to other dairy cattle breeds. Nonetheless, they have uniform milkability traits, and a large representation of desirable milk flow curves which are associated with a beneficial effect on the udder health.
The probability of contamination of non-transgenic varieties with genetically modified (GM) products increase as a result of global expansion of areas sown with transgenic crops. DNA-based methods as accurate, efficient and reliable methods are preferable for detection of GM material in raw or highly processed foods. Isolation of high quality DNA with a suitable and efficient DNA extraction protocol is crucial for getting precise results in DNA amplification. In this study, we performed modifications of previously known Sodium dodecyl sulfate (SDS)-based DNA extraction method regarding the incubation period, DNA pellet washing and addition of organic solvent extraction, to improve DNA quality and to reduce costs. Raw corn kernels and roasted soybean seed were used as samples. DNA was extracted following three protocols, modifications of Edwards protocol. The type of detergent used in raw corn sample did not cause significant effects on extracted DNA yield and purity, while in roasted soybean samples the 2% (w/v) SDS lysis buffer gave the highest DNA yield. The additional incubation step raised the DNA yield from raw corn for 121%, while the purest DNA from soybean sample was obtained using organic solvent extraction. Electrophoretic determination of DNA integrity showed varying degree of DNA smearing from roasted soybean. Contrary, all extraction protocols used on raw corn kernels produced a high molecular weight DNA. Thus, our in-house DNA extraction protocol is as efficient but more cost effective compared to commercial kits and can be used for raw corn, while the protocol for roasted soybean needs further improvement.
The aim of the research was to establish the morphology of the vascular bodies of the encephalon ventricles of cow (Bos taurus taurus). Methods used: thin anatomical preparation, histological method of examination, transmission electron microscopy. Given the relationship, structure and general origin of the vascular bodies, they were divided into bodies III, IV and the lateral ventricles of the encephalon. This unit has topographical nature. Functionally, vascular bodies are an indivisible organ whose main function is the secretion of the cerebrospinal fluid, which maintains the constancy of the central nervous system internal environment. Three types of capillaries, differing in their morphology, were found in the composition of the vascular bodies’ villi in Bos taurus taurus.
Subclinical mastitis is an asymptomatic udder infection distributed worldwide with enormous losses in the dairy industry. The study’s objective was to determine the presence of this pathological condition in small dairy farms in the R. of N. Macedonia and to identify the most common associated bacteria. Milk samples were obtained from 96 dairy cows (378 udder quarters) in seven dairy farms, in 3 consecutive samplings 24–72 hours apart. The samples were cultured on routine bacteriological growth media and incubated for 24–48 hours. The isolates were identified by AximaiD Plus MALDITOF MS Platform. Subclinical mastitis was found in 49 animals (51%) and 104 infected quarters (27%). The most frequent isolated bacteria on cow level were Streptococcus uberis (19.4%), Staphylococcus haemolyticus (13.4%), Staphylococcus aureus (7.4%) and Staphylococcu ssimulans (7.4%). On quarter level, the most isolated pathogen was Streptococcus uberis (35.6%) followed by Staphylococcu shaemolyticus and Staphylococcus aureus (10.3% and 9.2% respectively). Subclinical mastitis was found to be highly present in the selected small dairy farms. The most prevalent bacteria identified in the dairy farms (Streptococcus uberis, Staphylococcus aureus and coagulase–negative staphylococci) indicate that poor management and udder health practices, inadequate milking procedures and lack of mastitis control strategies greatly contribute to occurrence and persistence of subclinical mastitis.
This study provides insight into the development and validation of the simple, rapid and sensitive method of Liquid Chromatography-Tandem Mass Spectrometry for determining the residues of dexamethasone in bovine milk. The maximum residue limit for dexamethasone in bovine milk is 0.3 ng/ml−1. Chromatographic separation was achieved on an Agilent Proshell 120 SB:C18 (2.7µm 100x3.0 mm) column. The bovine milk was extracted with ethyl acetate, and the evaporated elution was dissolved with hexane, water and methanol, and analyzed by the LC-MS/MS method. The mobile phase A was a solution of 5 Mm ammonium formate in water, while the mobile phase B was 0.1% formic acid in acetonitrile. The calibration graphics were prepared within the range of 0.15–0.6 ng/ml−1, and a successful linearity was achieved (r ≥0.999). The limit of detection was 0.016 ng/ml−1. The decision limit (CCα) and detection capability (CCβ) were 0.34 and 0.38 ng/ml−1, respectively.