Serum anti-Müllerian hormone is an indirect predictor of ovarian reserve in domestic cats.

Anti-Müllerian hormone (AMH) serves as an indirect marker for predicting primordial follicles that are representative of ovarian reserve. In this study the possibility of using AMH and age to predict the ovarian reserve in domestic cats. Ovaries and blood were collected from 30 cats undergoing routine ovariohysterectomy. The animals were divided into three age groups: prepubertal (<4 mo, n = 10), adult (1-5 y, n = 10), and senior (>5 y, n = 10). Blood was collected at surgery for serum AMH measurements using the AMH Gen II ELISA kit. The intra-assay coefficient of variation (CV) and inter-assay CV were 3.56 % and 7.68 %, respectively. One side of the ovary was processed to determine AMH localization using immunohistochemistry and for a histological count of follicles, which is the gold standard. The expression of AMH protein was quantified from the contralateral ovary by Western blot analysis. Primordial follicles exhibited the most pronounced inverse relationship with age (rho = -0.779, P < 0.05), followed by a positive association with serum AMH concentration (rho = 0.490, P < 0.05), indicating that both age and AMH are potential markers indicative of primordial follicles. Furthermore, secondary (rho = 0.651, P < 0.05) and small antral follicles (rho = 0.648, P < 0.05) were identified as the major sources of circulating AMH, as indicated by the stronger correlation with serum AMH concentrations compared with primary follicles. However, there was no significant correlation between the expression of AMH protein and other factors, including age, primordial follicles, primary follicles, secondary follicles, small antral follicles, and serum AMH concentration. A model for predicting primordial follicle number using serum AMH concentration (AIC = 672.66, P < 0.05) and age (AIC = 668.93, P < 0.05) was established. In conclusion, both serum AMH concentration and age may serve as comparable markers of ovarian reserve in domestic cats. Moreover, AMH is particularly useful in situations where age information is not available.

Analysis of serum proteomic profiles of endangered Siamese and Burmese Eld's Deer infected with subclinical Babesia bovis in Thailand.

The endangered Eld's deer is a conserved species in Thailand, where tropical parasitic infections are endemic. Although Eld's deer with babesiosis are generally asymptomatic, they can still harbor the parasite and serve as reservoirs for ticks, spreading the infection to healthy animals within the herd. The present study aimed to investigate potential serum proteome biomarkers of Eld's deer with subclinical Babesia bovis infection. A total of 67 blood samples were collected from captive Siamese and Burmese Eld's deer showing no signs of parasitic infection. The nested polymerase chain reaction (nPCR) of a conserved spherical body protein 2 (sbp-2) gene of B. bovis was utilized to classify Eld's deer groups, with 25.37% (17/67) testing positive for B. bovis. Additionally, the application of proteomic studies showed that six B. bovis proteins, such as Obg-like ATPase 1 (OLA1) and heat shock protein 90 (HSP90), were significantly upregulated by more than a two-fold change compared with the PCR-negative samples. Of the 55 overexpressed serum proteins in the PCR-positives, alpha 2-HS glycoprotein (AHSG) and immunoglobulin lambda variable 2-8 (IGLV2-8) were notably among the top 10 proteins with the highest area under curve (AUC) values. Hence, they were proposed as potential biomarkers for subclinical B. bovis infection in Eld's deer. Analysis of the protein interaction network revealed interactions between Eld's deer AHSG and B. bovis OLA1 and HSP90, alongside associations with other proteins such as erb-b2 receptor tyrosine kinase 2 (ERBB2) and epidermal growth factor receptor (EGFR). These interactions were involved in the immune system pathway and inflammatory responses. Our findings shed light on subclinical infection of B. bovis in Eld's deer and identify potential biomarkers, contributing to the further effective detection and monitoring of B. bovis infection in this endangered species.

Biological Compositions of Canine Amniotic Membrane and Its Extracts and the Investigation of Corneal Wound Healing Efficacy In Vitro.

The usage of canine amniotic membrane (cAM) is mainly of interest in veterinary ophthalmology. Topical formulations of cAM could deliver the beneficial properties of cAM without the need for surgical intervention. The present study aimed to investigate biological compositions of cAM and its extracts, including their corneal wound healing efficacy. In this study, canine amniotic membrane extract (cAME) and lyophilized canine amniotic membrane extract (cAMX) were developed. Bioactive molecules related to corneal wound healing, including hepatocyte growth factor, tissue inhibitor of metalloproteinase-1 and -2, Thrombospondin-1 and Interleukin-1 receptor antagonist were studied at both gene and protein expression levels. Cell viability and wound healing assays were investigated for the possibility of cAME and cAMX as topical applications. The results demonstrated that all of the relevant genes and proteins were detected in cAM, cAME and cAMX. Both cAME and cAMX showed wound healing properties in vitro and cAME at 1.0 mg/mL concentration appeared to have the best healing efficacy. In conclusion, cAME and cAMX generated for topical use provided promising results in the healing of corneal defects.

Efficacy of bubaline blood derived fibrin glue in silk ligature-induced acute periodontitis in Wistar rats.

BACKGROUND AND AIM: Fibrin forms in the coagulation process, enhancing local hemostatic properties and promoting wound healing. The study aimed to evaluate the efficacy of bubaline-derived fibrin glue in silk ligature-induced periodontitis rats. MATERIALS AND METHODS: Bubaline blood-derived fibrin glue was prepared using cryoprecipitation and cryocentrifugation. Periodontitis was induced in rats by placing 5-0 silk ligatures around the mandibular first molars. The animals were divided into two groups: (1) Non-treatment and (2) bubaline fibrin glue-treated groups. Plaque, gingival inflammation, and mobility index were scored on days 1, 7, and 14 after intervention. Histological examinations were performed. The mRNA expression of inflammatory cytokines and growth factors was evaluated using a real-time polymerase chain reaction. Ligature-induced periodontitis was confirmed by the increase in inflammatory cell infiltration as well as histological bone and attachment loss. RESULTS: Compared to the non-treatment group, bubaline fibrin glue application reduced mononuclear cell infiltration into periodontal tissues corresponding to the reduction of collagen destruction. On days 7 and 14 after intervention, the inflammatory score and histological attachment loss were significantly lower in the bubaline fibrin glue-treated group than in the non-treatment group. A significant reduction in histological bone loss was observed in the treated group on day 7. Bubaline fibrin glue application led to a significant reduction of Tnfa and Il1b mRNA levels, while an increased expression of Pdgfa, Tgfb1, and Il10 was observed compared with the control. CONCLUSION: Bubaline fibrin glue could be beneficial in periodontitis treatment aiming to reduce inflammation and delay the progression of periodontal disease.

Serum protein profiles and C-reactive protein in natural canine filariasis.

BACKGROUND AND AIM: Canine filariasis is caused by several species of filarial worms. The pathophysiological response to infection is mainly due to the filaria lifecycle. Laboratory detection methods to assess the pathological alterations characteristic of filariasis are needed urgently. Serum protein profiles and C-reactive protein (CRP) levels are used widely to diagnose several animal diseases. This study aimed to determine the serum protein profiles and CRP levels in dogs infected with Dirofilaria immitis or Brugia pahangi or both parasites. MATERIALS AND METHODS: Blood samples were collected from 980 dogs presenting at animal hospitals and veterinary clinics in Bangkok and its vicinity. The presence of microfilaria in samples was determined using a buffy coat smear and staining with Wright-Giemsa. The sheathed and unsheathed microfilaria species were identified by acid phosphatase staining. Forty positive samples were tested. The serum protein profiles were identified by agarose gel electrophoresis. The CRP concentration was measured using a fluorescent immunoassay. RESULTS: Albumin levels and albumin-to-globulin ratios were significantly lower, and total protein, ß2 globulin, and γ globulin levels were significantly elevated in dogs infected with D. immitis and B. pahangi compared with reference values in normal dogs. The average CRP concentrations in dogs infected with D. immitis or B. pahangi were 69.9 and 12.9 mg/L, respectively. CONCLUSION: The total protein and γ globulin levels increased in canine filariasis compared with the normal reference range. The CRP concentration in dogs infected with D. immitis was extremely high, whereas that in dog infected with B. pahangi was normal.

Autoxidation of brain homogenates from various animals as measured by thiobarbituric acid assay.

INTRODUCTION: The TBARS assay has been well recognized for determination of lipid peroxidation and oxidative injury in biological samples including brain homogenates. In general, the homogenates are freshly prepared using rat brains as the tissue sources. In this study, we compared the rates of spontaneous lipid peroxidation in brain homogenates obtained from bovine, canine, hen, rat, and swine. In addition, the influences of lyophilization process and storage time up to six months at -20 degrees C without the freeze-thaw cycle were also determined in the swine brain preparations. METHODS: The standard assay for thiobarbituric acid-reactive substances (TBARS) was performed at 37 degrees C, using spectrophotometry to quantify the malondialdehyde (MDA) content. RESULTS: Rat brain homogenate exhibited the highest autoxidation rate (0.128+/-0.002 microM/min) whereas the bovine brain exhibited the lowest rate (0.032+/-0.001 microM/min). Swine brain homogenate could be kept at -20 degrees C up to 3 months without a significant increase in rate of autoxidation. Lyophilization caused a significant increase in the autoxidation rate of brain homogenate. However, the autoxidation rates of the lyophilized preparation were quite comparable throughout the six-month freezing time. DISCUSSION: Swine brain was a good candidate for tissue source in the TBARS reaction. The homogenate could be kept in the lyophilized form under the storage condition at -20 degrees C without the freeze-thaw cycle in the dark for at least six months.