Clonal spread of non-O157 Shiga toxigenic Escherichia coli O21:H25 in raw water buffalo milks.

AIMS: This study was conducted to investigate the presence of Shiga toxin-producing O157 and non-O157 E. coli in raw water buffalo milk, as well as to determine the virulence gene profiles, phylogroups, sequence types, and serotypes of the isolated strains. METHODS AND RESULTS: A total of 200 hand-milked raw water buffalo milk samples were collected from 200 different water buffaloes over a period of three months from 20 different farms. Isolation of STEC was performed using CHROMagar STEC. Presence of stx1, stx2, and eaeA genes were investigated by mPCR. Phylogroups and sequence types of E. coli strains were determined by Clermont phylotyping and MLST. Serotyping was performed using PCR or WGS. According to the results, two milk samples obtained from two different farms were found as STEC-positive. All Stx-positive E. coli isolates belonged to phylogenetic group A and were assigned to ST10. WGS results indicated that serotype of two isolates was O21:H25 and average nucleotide identity was detected at 99.99%. Thirteen additional registered E. coli O21:H25 assembled WGS data were obtained from EnteroBase and a phylogenetic tree was constructed. CONCLUSIONS: With this study, the presence of stx2 harboring E. coli O21:H25 in milk was identified for the first time. Although the identified serotype is considered a non-pathogen seropathotype, we conclude it could play an important role in the environmental circulation of Stx-phages and consequently contribute to the emergence of new STEC-related outbreaks.

Antioxidant and analgesic potential of butorphanol in dogs undergoing ovariohysterectomy.

The aim of this study was to evaluate the postoperative analgesic and antioxidant effects of butorphanol given in the preoperative or early postoperative period. Twenty-seven healthy female dogs undergoing ovariohysterectomy were randomly divided into three groups as before surgery group (BSG, n = 7) received butorphanol 30 min before preanesthetic administration, after surgery group (ASG, n = 10) received butorphanol during the last skin suture and the control group (CG, n = 10) received no butorphanol. Pain was assessed with short form of the Glasgow composite pain scale (CMPS-SF). Serum concentration of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase activities (GPx) were quantified by spectrophotometric methods to assess oxidative stress status. The pain score increased rapidly at 1 h after surgery and then decreased gradually towards to 24 h in all groups. There was no statistical difference among the groups in terms of CMPS-SF scores (P > 0.05). Serum concentration of MDA was lower in ASG than in BSG and CG from 1 h to 24 h after surgery. Serum activity of GPx was higher in ASG than in BSG and CG from 2 h to 24 h (P < 0.05). Serum activity of SOD was higher in ASG than in BSG and CG from 1 h to 24 h after surgery (P < 0.05). Serum SOD activity at different time points in ASG did not differ compared to preoperative level though it decreased significantly from 1 h onwards both in CT and BSG. The results indicate that single butorphanol administration either before or after the operation might not provide sufficient analgesia, however, it seems that it has antioxidant potential and may protect tissues by reducing oxidative stress when administered early postoperative period following ovariohysterectomy.

Expression of Anti-Müllerian Hormone and Its Type 2 Receptor in the Ovary of Pregnant and Cyclic Domestic Cats.

To evaluate the expression of AMH and its receptor AMHRII, ovaries of 33 p cats were investigated by western blot and immunohistochemistry. After ovariohysterectomy, the cats were grouped according to pregnancy stages and ovarian/placental endocrine activity: group I (n = 3, 24−29 days), II (n = 8, 32−40 days), III (n = 4, 41−46 days), IV (n = 6, 53−61 days) and according to cycle stages: V (n = 6, interestrus) and VI (n = 6, estrus). Serum progesterone- and AMH-concentration was measured. Follicle numbers did not differ between groups. The number of corpora lutea was higher in pregnant cats than in the non-pregnant cats. Serum AMH concentration was at maximum between day 30 and 50 of gestation, and was higher than in non-pregnant cats, then decreased towards term (p < 0.05). In the ovaries, AMH immunopositivity was observed in granulosa cells of secondary and antral follicles, and in interstitial cells of corpora lutea; highest percentage of immunopositive areas was detected in group III (p < 0.05). A positive correlation between the number of corpora lutea and the positive AMH signals in ovarian tissue was determined (r2 = 0.832, p < 0.05); however, only during mid-gestation (group II). Expression of AMHRII was in close co-localization with AMH and strong in the interstitial cells surrounding follicles undergoing atresia. AMHRII expression did not differ between pregnant groups but was higher compared to estrus cats (p ˂ 0.05). We conclude that AMH and AMHRII expression in the feline ovary is comparable to other species. The high serum AMH concentration and ovarian AMHRII expression between day 30 and 50 of gestation are probably related to ovarian activity and follicular atresia.

Effect of the stage of estrous cycle on follicular population, oocyte yield and quality, and biochemical composition of serum and follicular fluid in Anatolian water buffalo.

This study was designed to investigate the effects that the different stages of the estrous cycle had on the number of surface ovarian follicles and oocyte yield and quality of Anatolian water buffalo during peak breeding season. Assessments were made on the basis of ovarian morphology, serum and follicular fluid concentrations of variety of biochemical parameters. Following slaughter, blood samples were collected from each animal. The stage of estrous cycle was classified as either the luteal or follicular phase, and surface ovarian follicles were classified as small, medium, or large. The follicular fluid was aspirated, and oocytes were evaluated microscopically for classification into four categories. No statistical differences (p>0.05) were observed regarding the total number of follicles or quality of oocytes relative to the stage of the estrous cycle. Serum high-density lipoprotein cholesterol (HDL), alkaline phosphatase (ALP) and progesterone (P4) concentrations were significantly higher in the luteal phase than in the follicular phase (P<0.05). Significant correlations were observed in the luteal phase between the total number of oocytes and cholesterol (Cho), HDL, sodium (Na), chloride (Cl); A-quality oocytes and Na, Cl, Mg; C-quality oocytes and Cho, HDL, and Mg in follicular fluid. These results offer new information concerning Anatolian water buffalo reproductive physiology, which may be useful for improving oocyte quality in buffalo. This is the first study to describe the number of ovarian follicles, oocyte yield and quality, and a variety of biochemical parameters in the serum and follicular fluid of Anatolian water buffalo during peak breeding season in Turkey.