فصلنامه دامپزشکی ایران
سال بیستم شماره 82 (بهار 1403)

The aim of this research was to examine the serum vitamin D status in Gray Shirazi ewes during different reproductive stages, including mating, pregnancy, and early lactation. A total of 60 healthy ewes were sampled at three stages. The first stage was during mating, the second stage was in the late pregnancy, and the third stage was after delivery. The blood samples were analyzed for 25-hydroxy vitamin D levels. At delivery, multiple births, weight of lambs, and stillbirth rates were recorded. The results revealed that the mean serum vitamin D levels were significantly higher in the third stage of sampling (beginning of lactation) compared to the first stage (mating) and the second stage (pregnancy) (P=0.014 and P=0.017). However, there was no significant difference in vitamin D levels among the non-pregnant (the ewes that did not become pregnant after mating), singleton, and twin groups at any of the sampling stages. Vitamin D levels were highest at the beginning of lactation in gray shirazi ewes. Additionally, serum vitamin D was slightly decreased in pregnant ewes as the number of lambing increased. In total, the findings suggest that maintaining sufficient vitamin D levels may play a role in reproductive health in sheep, as there appears to be a correlation between vitamin D levels and fertility.

Polycystic ovary syndrome (PCOS) is one of the most important endocrine disorders affecting about 5-10% of women in reproductive age. Betaine is a bioactive peptide that has an anti-inflammatory activity. The present study aimed to evaluate the effects of betaine on insulin resistance indices and inflammatory cytokines markers and histology alteration in rat ovary with experimental PCOS. This experimental study was performed on 48 female Wistar rats (170-200 g). PCOS was induced by administration of testosterone enanthate (1 mg/100g/day during 35 days). PCOS rats were treated with betaine (1% in drinking water) for 30 days. At the end of the experimental period, the insulin resistance markers (serum insulin and glucose concentrations the homeostasis model assessment of basal insulin resistance (HOMA-IR), serum estradiol (E2), progesterone (P4) and inflammatory cytokines were measured. The ovaries were also processed for histological study. PCOS induction resulted in insulin resistance, impaired E2, P4 production and elevation of tissue TNF-α, IL-1β levels. Moreover, a significant increase of ovarian cysts and atretic follicles and also a significant reduction in the early corpus luteum were observed in the ovarian tissue of the PCOS groups. Betaine treatment could reduce the inflammatory cytokines and improve steroid productions in PCOS rats. Betaine also restored normal Folliculogenesis by reducing atretic and cystic follicles. Our findings showed the beneficial effects of betaine in PCOS rats by improvement of folliculogenesis, suppression of inflammation in ovary.

Sodium valproate (SV), as a common anti-epileptic drug, causes teratogenic effects on skeletal system by reactive oxygen species (ROS) generation. Herbal extract of Prosopis Farcta (PF), as a natural antioxidant necessary for many physiological activities, can probably ameliorate the teratogenic effects of SV during pregnancy. This study aimed to investigate the possible anti-teratogenic role of PF and Vitamin E (VE) on skeletal anomalies caused by SV in rat fetuses. Adult female rats (n=30) were categorized into 6 groups including control, SV (400 mg/kg), SV+VE (100mg/kg), and three doses of PF (50, 100, and 150 mg/kg) + SV. Each male rat mated with three adult female rats. The rats received SV, PF and VE at the 8th and 9th days of pregnancy by intraperitoneal injection. The animals were anesthetized and the laparotomy was applied at the 20th day of pregnancy. Skeletal abnormalities were analyzed using Alizarin red and Alcian blue staining. The expression of Runx2 and BMP2 genes was assessed using qPCRTM analysis in limbs bones. SV showed significant teratogenic effects including decrease in the rate of animal weight, Crown-rump length (CRL), various skeletal anomalies. The mRNA expression of Runx2 and BMP2 was also reduced in SV exposed animals. Administration of PF (especially 100mg/kg) in SV-exposed animals increased the weight of animals, CRL index, expression of Runx2 and BMP2, and reduced skeletal anomalies. The body weight, CRL index, Runx2 and BMP2 mRNA expression significantly increased, and skeletal anomalies decreased in VE group compared to the SV group. The results showed that PF could ameliorate the skeletal abnormalities and thus decreased osteogenic associated genes induced by SV in rat offsprings.

The present experiment was conducted to determine the chemical composition, digestibility and fermentation of red Alternanthera sessilis and compare its nutritional value with alfalfa either individually or in the form of rations of fattened lamb diets. Four experimental treatments included 1: control (without Alternanthera sessilis), 2: ration containing 3% (20% alfalfa substitution), 3: 6% (40% alfalfa substitution) and 4: 9% (60% alfalfa substitution) Alternanthera sessilis. Digestibility and fermentation of experimental samples were measured by gas production and two-step digestion test. In terms of chemical composition, there was a great relationship between Alternanthera sessilis and alfalfa hay. Adding Alternanthera sessilis to feed of lambs significantly increased the amount of truly degraded organic matter, metabolizable energy, and apparent digestibility of organic matter, so that these parameters were highest in treatment containing 9% of Alternanthera sessilis plant. On the other hand, by using Alternanthera sessilis plant, dry matter digestibility tended to increased. The amount of microbial biomass production and microbial biomass production efficiency were not affected by experimental diets. Rations containing Alternanthera sessilis were cheaper than control diets. Therefore, it can be stated that the nutritional value of Alternanthera sessilis is comparable to alfalfa hay and can be replaced by alfalfa by up to 60% in the ration of fattening lambs. In vivo experiments are also suggested.

Myocardial infarction (MI) has increased in the recent years due to aging as well as lifestyle changes in the population. In the last decades, regenerative medicine has been considered to provide modern and efficient methods for MI patients' therapies. The current study was aimed at investigating the effect of acellular aortic scaffold (AAS) on MI cardiomyocytes to identify the therapeutic potential of this natural biomaterial for MI patients. The study was conducted in five main steps: Preparing MI animal model, preparing MI cells, culture of animal MI cell on AAS, assessment of cell viability (MTT assay) as well as antioxidant and catalytic activities (ROS, TAC, SOD, GPX, and CAT assays) and molecular study on apoptotic and anti-apoptotic factors (qRT-PCR). AAS positively affected the viability rate of MI cells and the GPX and SOD levels were significantly increased in MI cells due to the culture on AAS. The RT-PCR quantification showed a decrease in the levels of of Cox8 and Caspase3 expression genes levels in MI cells, cultured on AAS, while an increase in miR-24 expression level was observed in MI cells as a result of culture on AAS. We concluded that acellular aortic scaffold can positively control the catalytic and antioxidant activities, cellular hemostasis, and cell viability of cardiomyocytes after myocardial infarction, demonstrating the potential of such natural biomaterials for cardiac tissue reconstruction. However, to achieve favorable results and ideal therapeutic applications, further studies are required.

Agricultural by-products are frequently used to feed livestock, but they often have low nutritional value. This study was conducted to improve nutritional value of wheat straw using cellulolytic bacteria isolated from the foregut of dromedary camel. Rumen fluid was collected from four fistulated camels. The cellulose-hydrolytic bacteria were isolated by using medium amended with carboxymethyl cellulose (CMC). The activity of carboxymethyl cellulase (CMCase) was determined by measuring the release of reducing sugars. The optimum pH and temperature for growth and CMCase activity of the isolates were determined. Three isolates (SH13, SH2, and SH22) showed maximum hydrolysis capacity and were selected for further investigation. The analysis of 16S rRNA gene sequences of these isolates showed that the isolates possessed 99, 94 and 98% similarity with Escherichia coli, and Enterobacteriaceae, respectively. Wheat straw was incubated with the isolated bacteria in liquid medium for 6 weeks. The results showed inoculation with SH22 isolates resulted in the highest disappearance of dry matter and crude protein content, and the lowest amount of neutral detergent fibre (NDF), iNDF and acid detergent fibre (ADF). Treatment with SH13 isolate had the highest digestibility of NDF and ADF, and treatment with SH2 had the highest truly degraded organic matter (TDOM) and partitioning factor (PF). Our data suggest that maybe bacteria isolated from camel foregut can increase digestibility and enhancing nutrition value of wheat straw.

Synbiotics, which are a combination of prebiotic and probiotic supplements, are one of the most commonly used feed additives in aquaculture. Present study was carried out to evaluate the effect of dietary supplement of two strains of intestinal bacteria Lactobacillus plantarum and Lactobacillus pentosus from the intestinal tract of the Shabot (Tor grypus) with β-glucan on growth performance and digestive enzyme activity of rainbow trout (Oncorhynchus mykiss) fingerlings. The fish were divided randomly into 8 treatment groups (for 60 days) containing control (group 1), 1% β-glucan (group 2), Lactobacillus plantarum (group 3), Lactobacillus pentosus (group 4), Lactobacillus plantarum + Lactobacillus pentosus (group 5), Lactobacillus plantarum with 1% β-glucan (group 6), Lactobacillus pentosus with 1% β-glucan (group 7), Lactobacillus plantarum + Lactobacillus pentosus with 1% β-glucan (group 8). After 60 days, the fish fed combination of two probiotics at equal proportion with 1% β-1,3-glucan (group 8) had higher growth performances than the other treatment groups. Digestive enzyme activities such as ALP, α-amylase, trypsin, lipase and protease in groups including probiotics and prebiotic, particularly in group 8 in parallel with growth performances, had higher than other groups. However, the enzyme activity of chemotrypsin had no significant difference between treatments. These results indicate that a combination of host-derived probiotics (L. plantarum and L. pentosus) with β-1,3-glucan has a significant potential as an important synbiotics to enhance the nutrients utilization and activity of digestive enzymes in rainbow trout juveniles; However, further research is needed to determine the proper supplement for this commercially valuable product.

Lactobacillus probiotic enriched diets can modulate host immune response. In this study, we investigated whether the three species Lactobacillus plantarum, L. bulgaricus and L. rhamnosus up-regulated two T-cells specific antigens and exerted the protective effect when administrated with formalin-killed Streptococcus agalactiae in Nile tilapia. For this purpose, a total of 180 Nile tilapia (average body weight 45.8 ± 22 g) were randomly divided into 6 experimental groups, i.e.: one control group and 5 vaccinated groups: Formalin killed cell (FKC); FKC+ Adjuvant; FKC+ L. plantarum; FKC+ L. bulgaricus and FKC+ L. rhamnosus. All groups were fed with normal commercial pellets and three groups were fed with pellets sprayed with three different lactobacilli. The results showed that the survival rate in the groups of Formalin killed cell (FKC) combined with oral administration of lactobacilli was from 70.0% to 75.0%.  Protection in the control group amounted to 31.3% illustrating a significant difference with other experimental groups. In this research, gene expression of CD4 and CD8 which have essential functions in the immune response quantified by qRT-PCR in the head kidney, skin, and spleen was reported in the form of fold change. The analysis of fold change (Mean ± SD) related to CD4 and CD8 at 30- and 60-days post-immunization (dpi) respectively showed an increase in fold change in all probiotic groups compared to the FKC group in two of which it was significant at the level of p<0.05. The results revealed that vaccination with FKC administrated lactobacilli-enriched diets increased the expression of genes related to immune response which can indicate higher protection against Streptococcus agalactiae in the probiotic groups compared with FKC vaccination alone similar to the results observed in FKC coupled with adjuvant vaccination.

Menace response and pupillary light reflex are used as part of the ophthalmic and neurological examinations in animals. This study was conducted to investigate the menace response and pupillary light reflex in native calves that were not separated from their mothers after birth and had traditional rearing conditions. Thirty healthy native calves were included in the study. The menace response was assessed by applying a non-tactile visual threat to each eye. The direct pupillary light reflex was tested by shining a bright light into each eye and observing constriction of the pupil in the ipsilateral eye. Calves were observed after birth until the menace response and pupillary light reflex were well present. The pupillary light reflex was positive in all calves in the first test performed after birth. The menace response developed in the age range of 3 to 6 days in calves and this response developed earlier in male than female calves. According to the results of previous study, none of the calves developed a menace response during the time they were housed in individual stalls. The present study indicated that development of menace response in traditionally reared calves (housing with their mothers in a common pen) was observed on the 6th day after birth.

It has been determined that there is a direct relationship between the severity of mastitis and the virulence factors produced by bacterial agents. Identifying bacterial virulence factors is neccecary for designing suitable vaccines against mastitis. The aim of the present study was molecular diagnosis of selected virulence factors of endemic isolates of S. aureus involved in bovine mastitis. A total of 180 milk samples were collected from cows with clinical (37 samples, 20.6%) and subclinical (143 samples, 79.4%) mastitis from 8 semi-industrial dairy farms of Chaharmahal and Bakhtiari province, Iran. After culture and purification, coagulase, catalase and oxidase tests were performed. DNA was extracted from S. aureus suspected colonies. Final confirmation was performed using PCR test on the specific 23S rRNA gene of the bacteria. Thirty one (17.22%) out of the 180 collected samples were found to be positive for S. aureus by PCR, of which 2 cases were related to clinical mastitis and 29 cases were related to subclinical mastitis. The highest frequency of virulence genes was related to the Coa gene (90.32%), followed by ClfB (87.09%), LukD, and fnbB (80.64%), LukE (77.41%), fnbA (74.19%), Hla (48.38%). The lowest frequency was related to the Hlb gene (45.16%). Based on the obtained results, the diagnosis of the virulence factors of S. aureus has the potential of being used in the development of vaccines for the prevention of mastitis.

The scorpion of Androctinus crassicauda is one of the Buthida family members whose neurotoxic venom is deadly to humans. Enzymes are abundant components of animal venom that play roles in local and systemic symptoms following envenoming. The venom of this scorpions contains various pathological enzymes, among which studies have reported the toxic effects of venom's hyaluronidase and phospholipase A2 enzymes. This study aimed to measure the activity of venom enzymes of hyaluronidase and phospholipase A2 from scorpion of A. crassicauda and its fractions. For this purpose, the venom from scorpion of A. crassicauda were collected and lyophilized. The whole venom was fractionated on a gel filtration (Sephadex G-50) column. Then protein concentration, hyaluronidase and phospholipase A2 activity of the crude venom and its fractions were determined. Gel filtration on Sephadex G-50 and SDS-PAGE electrophoresis of A. crassicauda crude venom revealed four protein picks and 15 protein bands with the majority of molecular masses between 10.0 and 15.0 kDa, respectively. The supernatant obtained from crude venom clarification showed protein content 71.80%, high hyaluronidase (92.40%) and low PLA2 (302±2.1 mg/U) enzyme activity. Fraction 1 showed highest hyaluronidase and phospholipase A2 activities. We found that pH 5 is the optimum pH for hyaluronidase activity and phospholipase A2 activity increases with increasing incubation time. In conclusion, venom of A. crassicauda displayed the hyaluronidase and phospholipase A2 enzymes activity, which were separated by gel filtration chromatography in fraction I. Such information is important both for predicting the biological activity of venom components and useful for developing effective antivenoms based on these venom components.

Insect bite hypersensitivity is a recurrent and chronic dermatitis in horses, that is mainly caused by an allergic reaction to the bites of Culicoids species, and therefore in areas where these insects can grow, the risk of getting the disease also increases. The conditions in Khuzestan province (located in southwestern of Iran) are mostly hot and humid, causing many veterinarians to encounter horses with IBH symptoms. The present study aimed to evaluate the frequency of insect bite hypersensitivity among horses in this province. The study was performed on 255 Arabian horses kept in 21 horse breeding centers located in 6 cities of this province. The horses were examined for the presence or absence and location of skin lesions, along with the individual animal information, which was recorded in relevant forms. The prepared serum samples (32 healthy horses and 61 affected horses) were evaluated for total IgE using the ELISA test according to the instructions of the kit manufacturer. The results showed that 144 (56.5%) out of totally 255 horses had clinical signs of IBH. Statistical evaluations also revealed that among the hosting factors (including age, gender, bloodline, coat color, body condition score, and type of use), and management and environmental factors (including stud size, methods of control of insect, distance from the horse breeding centers to the water source, times of fecal collection in the horse breeding centers and the geographical location of the horse breeding centers), only stud size had a significant effect on the prevalence of IBH, In other cases, there was no significant relationship between the prevalence of the disease and the evaluated factor. The results also showed that the amount of IgE in affected horses was higher than in healthy horses. The results of this study showed that insect bite hypersensitivity exists with a significant frequency among horses kept in the horse breeding centers in Khuzestan province.