Milk progesterone enzyme-linked immunosorbent assay as a tool to investigate ovarian cyclicity of water buffaloes in relation to body condition score and milk production

Background

Application of assisted reproductive technologies in buffaloes is limited to some extent by farmers’ inability to detect oestrus because of its poor expression. The present study aimed at investigating reliability of a milk progesterone enzyme-linked immunosorbent assay (ELISA) to assess the ovarian cyclicity during post partum, oestrus and post-breeding periods in water buffaloes.

Methods

Progesterone concentrations were measured by an ELISA in milk of 23 postpartum buffaloes in relation to oestrus, pregnancy, body condition score (BCS) and milk production. Two milk samples were taken at 10 days intervals, every month starting from day 30 and continued to day 150 post partum. BCS and milk production were recorded during sample collection. Milk samples from bred buffaloes were collected at Day 0 (day of breeding), Days 10–12 and Days 22–24. Defatted milk was preserved at −80°C until analysis. Pregnancy was confirmed by palpation per rectum on Days 70–90.

Results

Seventeen buffaloes had 47 ovulatory cycles, one to four in each, 13 were detected in oestrus once (28 % oestrus detection rate). Progesterone concentration ≥1 ng/ml in one of the two 10-day-interval milk samples reflected ovulation and corpus luteum formation. The intervals between calving to first luteal activity and to first detected oestrus varied from 41 to 123 days (n = 17) and 83 to 135 (n = 13) days, respectively. Eight buffaloes were bred in the course of the study and seven were found pregnant. These buffaloes had a progesterone profile of low (<1 ng/ml), high (≥ 1 ng/ml) and high (≥ 1 ng/ml) on Day 0, Days 10–12 and Days 22–24, respectively. Buffaloes cycling later in the postpartum period had fewer missed oestruses (P < 0.05). Buffaloes with a superior BCS had a shorter calving to oestrus interval and produced more milk (P < 0.05).

Conclusions

Milk progesterone ELISA is a reliable tool for monitoring ovarian cyclicity and good BCS may be an indicator of resuming cyclicity in water buffalo.     

Rumen development and growth of Balouchi lambs offered alfalfa hay pre- and post-weaning

The consumption of solid feed is essential for successful transition from a pre-ruminant to a functional digestive tract. Lambs fed starter rations containing highly fermentable carbohydrates often experience dramatic changes in concentrations of rumen and blood metabolites. The optimal amount of roughage required in the diet of pre-ruminant animals is still unclear. The objective of this study was to determine the effect of feeding alfalfa hay on performance and rumen development in young Balouchi lambs. In a completely randomized design, 30 lambs were fed one of three experimental diets consisting of a control, without alfalfa hay (C), a diet containing 7.5% alfalfa hay (A1), and a diet containing 15% alfalfa hay (A2). Lambs fed A1 and A2 diets had lower dry matter intake during the pre-weaning period (P?P?=?0.02), but feed conversion ratio and average daily gain were not affected by feeding alfalfa hay. Concentration of beta-hydroxybutyric acid was higher in C compared with the A1 and A2 groups (P?P?=?0.04) and increased thickness of muscular layer (P?=?0.05). We concluded that including 15% alfalfa hay in the starter diet could reduce thickness of the keratinization layer and increase muscularity of rumen wall without adverse effects on growth and performance of newborn lambs.     

DNA vaccine-encoded glycoprotein B of HSV-1 fails to protect chronic morphine-treated mice against HSV-1 challenge

The use of morphine has been demonstrated to increase susceptibility to infections. Herpes simplex virus type 1 (HSV-1) is a highly successful pathogen among immunocompromised individuals. In the present study, due to the importance of HSV vaccination in morphine abusers, the effects of chronic morphine exposure on the host response to a HSV-1 gB DNA-based vaccine have been investigated. The study is addressing an important aspect of vaccine development among the susceptible (immunocompromised) hosts. BALB/c mice were exposed to morphine over 11 days. They were then vaccinated with DNA vaccine or KOS strain as a live vaccine. The findings showed that the morphine-treated animals failed to respond to DNA vaccination evaluated by the anti-HSV gB antibody titer, delayed type hypersensitivity (DTH) and lethal HSV-1 challenge. Under the same conditions, the KOS vaccine showed a reduced Ab titer and DTH response in morphine-treated mice, but could protect mice against the lethal challenge and was safe for vaccination of morphine-treated animals.     

Molecular typing of Iranian field isolates Mycoplasma synoviae and their differentiation from the live commercial vaccine strain MS-H using vlhA gene

1. The single-copy domain of the N-terminal region of the vlhA gene of Mycoplasma synoviae was sequenced, analysed and verified and used to type Iranian field isolates of M. synoviae and the MS-H live vaccine strain. In addition, a restriction fragment length polymorphism (RFLP) method was developed to differentiate between field isolates of Iranian and MS-H vaccine strains.

2. All sequences were analysed and aligned; the percentage similarity of the DNA was calculated and dendrograms were constructed. Based on single nucleotide polymorphism (SNP) that existed in all field isolates in Iran, the PCR-RFLP method allowed the differentiation of all M. synoviae field isolates from the vaccine strain.

3. Using phylogenetic analysis, the isolates were assigned to 8 unique genotypes and, within each group, DNA had a high level of similarity.

4. DNA sequence analysis and PCR-RFLP of the amplicon based on percent similarity and evolutionary relationship appeared to be useful tools for strain differentiation whether M. synoviae clinical isolates from infected chickens were derived from the vaccine strain or wild-type strains.

5. This study confirms the potential value of strain typing for epidemiological purposes and suggests that phylogenetic studies are essential to understand the true relationships between strains.     

Glutathione content of in vivo and in vitro matured canine oocytes collected from different reproductive stages

Glutathione (GSH) concentrations of oocytes are considered as an important marker of the cytoplasmic maturation. The present study was designed to compare GSH concentrations of in vivo and in vitro matured canine oocytes. In vivo matured oocytes were collected 72 hr after ovulation by flushing fallopian tubes after laparotomy. Ovaries were collected from bitches with different reproductive stages, and collected oocytes were divided into 2 groups according to the size viz. < 120 microm and > 120 microm in diameter and cultured for 72 hr in Tissue Culture Medium-199 supplemented with 10% FBS, 2.2 mg/ml sodium bicarbonate, 2.0 microg/ml estrogen, 0.5 microg/ml FSH, 0.03 IU/ml hCG, and 1% penicillin-streptomycin solution in the presence or absence of 50 microM beta-mercaptoethanol. GSH concentrations were determined by the dithionitrobenzoic acid-glutathione disulfide (DTNB-GSSG) reductase recycling assay. GSH concentrations of immature canine oocytes were 2.9 and 3.8, 3.5 and 6.8, and 3.1 and 6.5 pM/oocyte for < 120 microm and > 120 microm in diameter oocyte groups at anestrous, follicular and luteal stage, respectively (P<0.05). In vivo matured oocytes had significantly higher GSH concentrations compared with in vitro matured oocytes. The GSH content was 19.2 pM/oocyte for in vivo matured oocytes, while 4.1 to 8.1 and 5.7 to 13.2 pM/oocyte for in vitro matured oocytes cultured in the absence or presence of beta-mercaptoethanol, respectively (P<0.05). Presence of beta-mercaptoethanol increased GSH synthesis in canine oocytes cultured in vitro, and oocytes collected from follicular and luteal stage was superior to anestrus oocytes.     

Sero-prevalence of avian influenza among broiler-breeder flocks in Jordan

Thirty blood samples were collected randomly from each of the 38 breeder-broiler farms in Jordan. Serum samples were examined using indirect ELISA for specific antibodies to avian influenza virus. The overall true flock-level sero-prevalence of avian influenza was 71% (95% CI: 55,83). Positive flocks had 2-30 sero-positive chickens and half of flocks had >20 sero-positive birds. The number of sero-positive flocks varied in the studied localities with more sero-positives in farms located within the migratory route of migratory wild fowl. The examined broiler-breeder flocks had no clinical signs, or noticeable decrease in egg production; mortalities were within the normal range (0.1-1%). The number of positive sera/flock correlated with flock size. There were a no significant (Pearsons r=0.21, p=0.21) correlation between positive flocks and age. A non-pathogenic AI virus infects broiler-breeder farms in Jordan. Wild local and migrating birds might promote the further spread of this virus in Jordan and other countries.     

Occurrence of anti-<Emphasis Type="Italic">Toxoplasma gondii</Emphasis> antibodies in female cattle in south-west of Iran

Toxoplasmosis, caused by Toxoplasma gondii, is a significant disease in livestock and humans. In Iran, studies shows that T. gondii infection in humans is relatively high and prevalence is associated mainly with consumption of undercooked meat or meat products. We have examined 450 serum samples from female cattle distributed over all Ahvaz, the center of Khouzestan province, south-west of Iran. IgG antibodies to T. gondii were assayed by the modified agglutination test using whole tachyzoites of T. gondii, and found in 71 (15.77%) of 450 cattle with titers of 1:25 in 38, 1:50 in 18, 1:100 in 11, 1:200 in three and 1:400 in one. Titers of antibodies were decreased in cattle over 2 years old. These results indicate that T. gondii infection in cattle of Khouzestan is relatively considerable, but not very high and consumption of beef may be a source of infection for humans in south-west of Iran.     

Evaluating the correlation between adrenal gland dimensions and aortic diameter in healthy dogs

In the present study, the relationship of normal adrenal dimensions with weight, aortic diameter, age and sex was evaluated. Moreover, a formula for estimating the normal dimensions of adrenal gland was provided. Thirty‐two intact adult dogs that were considered healthy based on history, physical examination, routine blood works and specific adrenal tests as well as absence of any abnormal ultrasonographic findings were included. Adrenal glands and abdominal aorta were imaged in the sagittal plane by ultrasound. Length, maximal height of the cranial and caudal poles, area and circumference of the left and right adrenal glands and also the internal diameter of the abdominal aorta were measured. Analysis showed that there is a positive correlation between the adrenal gland dimensions and weight and aortic diameter. The ratio of adrenal gland dimensions and the aortic diameter was calculated, but this ratio showed a great amount of variability in tandem with a significant correlation to the weight; Meaning that the ratio of the adrenal gland dimensions to the aortic diameter was not a reliable index for evaluating the size of the normal adrenal gland in animals with different weights. Therefore, we used the weight along with adrenal measurements in a linear regression model, and then, we were able to estimate the adrenal gland dimensions in different weights. Knowing the relationship of normal adrenal dimensions with the indices such as weight, age, sex and diameter of abdominal aorta can help the clinicians to diagnose the adrenal gland diseases.     

Livestock feed resources availability,feeding practices,and nutrient balances in high Himalaya (Gurez) valley of Kashmir

Tropical Animal Health and Production - Assessment of the ruminant livestock feed sources availability, existing feeding practices, and nutrient balances was embarked at different zones of Gurez...