Cell-Specific Expression Pattern of Toll-Like Receptors and Their Roles in Animal Reproduction.

Toll-like receptors (TLRs), a part of the innate immune system, have critical roles in protection against infections and involve in basic pathology and physiology. Secreted molecules from the body or pathogens could be a ligand for induction of the TLR system. There are many immune and non-immune types of cells that express at a least single TLR on their surface or cytoplasm. Those cells may be a player in a defense system or in the physiological regulation mechanisms. Reproductive tract and organs contain different types of cells that have essential functions such as hormone production, providing an environment for embryo/fetus, germ cell production, etc. Although lower parts of reproductive organs are in a relationship with outsider contaminants (bacteria, viruses, etc.), upper parts should be sterile to provide a healthy pregnancy and germ cell production. In those areas, TLRs bear controller or regulator roles. In this chapter, we will provide current information about physiological functions of TLR in the cells of the reproductive organs and tract, and especially about their roles in follicle selection, maturation, follicular atresia, ovulation, corpus luteum (CL) formation and regression, establishment and maintenance of pregnancy, sperm production, maturation, capacitation as well as the relationship between TLR polymorphism and reproduction in domestic animals. We will also discuss pathogen-associated molecular patterns (PAMPs)-induced TLRs that involve in reproductive inflammation/pathology.

Altered luteal expression patterns of genomic and non-genomic progesterone receptors in bitches at different reproductive states.

The binding of steroid hormones to their specific receptors is necessary to exert their effects on target cells. Progesterone (P4), a steroid hormone, carries out its effects through both genomic and non-genomic (the cell membrane-associated) receptors. This study aimed to ascertain luteal expression patterns of genomic and non-genomic progesterone receptors in bitches in physiological (early dioestrus and early pregnant) and pathological (pyometra) reproductive states. Luteal tissue was collected from the bitches at early dioestrus (ED, n = 5), early pregnant (EP, n = 5), and pyometra (PY, n = 5). The expression profiles of Steroidogenic Acute Regulator Protein (STAR), Progesterone Receptor (PGR), Membrane Progestin Receptors (PAQR5, PAQR7 and PAQR8), and Progesterone Membrane Components (PGMRC1 and PGMRC2) were examined at the mRNA levels using Real-Time Polymerase Chain Reaction (RT-PCR). Protein levels of PGR, PGMRC1 and PGMRC2 were detected by western blotting (WB). The STAR expression was found in all groups, with a statistical difference observed between EP and PY groups (P < 0.05). The protein level of PGR was determined to be highest in the EP group and lowest in the PY group. The expression of PAQR8 increased in the EP group (P < 0.05). The PAQR5 exhibited high expression in the EP group and low expression in the PY group (P < 0.05). PGRMC1 was more elevated in the EP group and lower in the PY group (P < 0.05). Protein levels of PGMRC1 and PGMRC2 were also observed at the highest expression in EP group. According to the altered expression profiles for examined receptors, we suggest that those progesterone receptors have roles in early pregnancy or pyometra in bitches.

Salmonella, Campylobacter, and Chlamydophila in bald ibis (Geronticus eremita) feces in Turkey.

The aim of this study was to investigate the presence of Campylobacter spp., Salmonella spp., and Chlamydophila psittaci in fecal samples of bald ibises (Geronticus eremita) housed in a conservation facility in Turkey. A total of 82 fecal samples were collected from cages and evaluated by bacteriologic methods and a polymerase chain reaction (PCR) technique for Campylobacter spp. and Salmonella spp. and by PCR for C. psittaci. Campylobacter spp. were isolated from 24 of 82 fecal samples (29.2%). Of these 18 (75%), 4 (16.7%) and 2 (8.3%) were Campylobacter jejuni, Campylobacter coli, and other Campylobacter spp., respectively. Salmonella spp. were detected in 8 fecal specimens.(9.7%) by PCR. The presence of C. psittaci was not detected in the bald ibises studied. The results suggested that the bald ibises in this present study might be at a higher risk of infection with Salmonella spp. and Campylobacter spp.

Economic Importance of Hydatid Cyst in Slaughtered Cattle of Mugla Province

Objective: This study was carried out to investigate the prevalence, seasonal distribution and economic effects of hydatid cyst infection in slaughtered cattle of Mugla province. Methods: Data from the 2019 records of the Mugla Metropolitan Municipal Abattoir were studied retrospectively. Both direct and indirect calculation methods were used to determine the economic losses incurred due to hydatid cyst. Results: Twenty-one out of 9.985 (0.21%) cattle were found to have been infected with hydatid cyst in 2019. The highest prevelance of infection was reported in February (1.17%), while no cases were observed in May, June and August. The direct economic loss attributed to liver destruction in Mugla province was 11,760 TL (1.950 $). The total indirect economic loss caused by hydatid cyst (carcass loss, milk production loss, decreased fecundity) was 122,691 TL (20,346 $). Cumulatively, the direct and indirect economic losses for hydatid cyst disease in Mugla province were 134,451 TL (22,296 $). Conclusion: According to our results, the prevalence of hydatid cyst in cattle of Mugla province was 0.21%, which was still lower than the average loss in Turkey in spite of the corresponding significant economic loss.

Sequence diversity of MHC class-II DRB gene in gazelles (Gazella subgutturosa) raised in Sanliurfa of Turkey.

In this study, we aimed to assess the sequence diversity of major histocompatibility complex (MHC) class-II DRB gene at exon 2 in gazelles raised in Sanliurfa Province of Turkey. Twenty DNA samples isolated from gazelles (Gazella subgutturosa) were used for sequencing exon 2 of MHC class-II DRB gene. Target region was amplified by polymerase chain reaction (PCR) and their products were directly sequenced. Nine of these 20 samples yielded unambiguously readable sequences. Three of the nine samples were homozygotes and each showed different sequences. A 262-bp sequence obtained from the three homozygote samples were submitted to GenBank (accession numbers: KC309405, KC309406 and KC309407). Using an allele specific PCR, we detected 10 additional haplotypes. Among 13 haplotypes, 45 nucleotide positions were polymorphic and most of the polymorphic nucleotide positions localized at peptide-binding region (PBR). Rates of nonsynonymous substitutions were significantly higher than synonymous substitutions at PBR. Phylogenetic analysis of the haplotypes showed that 10 haplotypes of the gazelles were clustered together while three were clustered with ovine and bovine haplotypes. The results indicated that at least 13 haplotypes at exon 2 of MHC class-II DRB gene were showing high degree of nucleotide and amino acid diversity, and certain haplotypes of G. subgutturosa were more similar to haplotypes from sheep or cattle than to each other. Rates of synonymous and nonsynonymous substitutions suggested that positive selection was a driving force for diversity at this locus in G. subgutturosa.

The scintigraphic, biodistribution and histopathological evaluation of the effect of experimental 131I administration on the gastrointestinal system and the demonstration of the Na+/I- symporter by immunohistochemistry in rats.

OBJECTIVE: The aim of this study was to investigate histopathological changes and biodistribution of iodine-131 (I) in the gastrointestinal system (GIS) and also Na/I Symporter (NIS) presence by immunohistochemically in the experimental treatment of rats with radioactive iodine (RAI). MATERIALS AND METHODS: Rats were divided into experimental and control groups as random early group 2 (24 h), intermediate group 3 (3 weeks), and late period group 4 (3 months). Experimental groups were administered 100 MBq (∼3 mCi, 12 mCi/kg) by orogastric route with orogastric tube. Scintigraphic iodine screening images were obtained 24 h, 3 weeks, and 3 months after RAI, and GIS tissues were removed, and immunohistochemical methods were used to demonstrate NIS with RAI biodistribution and histopathology. RESULTS: According to the results of scintigraphy, the most prominent activity involvement was observed in the thyroid gland at group 2, and significant activity was observed in the stomach. In the group 3 and group 4 images, owing to the physiological and biological half-life of the iodine and low resolution of the gamma camera, no secondary focal activation was observed. The highest RAI biodistribution value in all groups was in the stomach, ileum and oesophagus. In the immunohistochemical examination of NIS, the highest staining sequence was observed in all groups respectively in the stomach, oesophagus, tongue, colon, saliva, duodenum, rectum, ileum and jejunum. The increase of NIS immunohistochemically stained more intensely was observed in the RAI-administered groups. CONCLUSION: The amount of NIS is important for the absorption of RAI after administration.

Expression patterns of Toll-like receptors in the ovine corpus luteum during the early pregnancy and prostaglandin F2α-induced luteolysis.

The aim of this study was to elucidate the expression profiles of Toll-like receptors (TLRs) in the ovine corpus luteum (CL) during early pregnancy and prostaglandin F2α (PGF2α)-induced luteolysis. For this purpose, multiparous Anatolian Merino ewes were selected and randomly allotted into cyclic (including those in the induced luteolysis group, n = 20) and pregnant (n = 12) groups. All of the ewes were scheduled to be slaughtered for predetermined days/hour during the estrous cycle, early pregnancy, and PGF2α induced luteolysis. The CLs were collected from both cyclic and pregnant ewes on days 12 (C12 and P12; n = 8) and 16 (C16 and P16; n = 8) and pregnant ewes on day 22 (P22; n = 4). For the induced luteolysis model, ewes were injected with PGF2α on day 12 of the estrous cycle and CLs were collected at 1 h (PG1h; n = 4), 4 h (PG4h; n = 4), and 16 h (PG16h, n = 4) after injection. Quantitative polymerase chain reaction (qPCR) was used to evaluate the expression profiles of TLR2, TLR4, TLR6, TLR8, and TLR10, while free-floating in situ hybridization and immunohistochemistry were used to define the spatial localization of TLR2, TLR4, and TLR7 in the CL. Data were then analyzed by one-way ANOVA and were considered statistically significant when P values were lower than 0.05. Expression of TLR2 was upregulated in both early and late stages of luteolysis (P < .05). An upregulation of TLR4 was detected at PG16h, while TLR6 was decreased at PG4h (P < .05). Expression of TLR7 and TLR8 was significantly increased during early pregnancy, at both PG16h and regressed groups (C16, P < .05). In contrast, TLR10 was downregulated during PGF2α-induced luteolysis and on P16 (P < .05). TLR4 and TLR7 proteins were particularly localized in endothelial cells on C12/PG0h, but prominent signals corresponding to TLR4 and TLR7 were detected in luteal cells at PG16h. The results suggest an involvement of TLRs in the luteolytic mechanism in ovine CL, as indicated by differential expression levels of TLRs during PGF2α-induced luteolysis. Moreover, the present study indicates that early pregnancy-mediated changes in TLR expression in the CL may contribute to the establishment and maintenance of ovine pregnancy.

Rating of putative housekeeping genes for quantitative gene expression analysis in cyclic and early pregnant equine endometrium.

The aim was an evaluation of a set of housekeeping genes (HKGs) to be used in the normalization of gene expression in the equine endometrium. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine ribosyl transferase 1 (HPRT1), ubiquitin B (UBB), tubulin alpha 1 (TUBA1), ribosomal protein L32 (RPL32), beta-2-microglobulin (B2M), 18S rRNA (18S), and 28S rRNA (28S) HKGs were evaluated using real-time PCR and were compared in different physiological stages of the endometrium. Endometrial biopsies were obtained from mares on day of ovulation (d0, n=4), at late diestrus (LD, n=4), after luteolyis (AL, n=4) of the cycle and on days 14 (P14; n=3), 18 (P18, n=3) and 22 (P22; n=3) of pregnancy. A model based on REML with support of descriptive statistics was proposed in accordance with experimental design and was further confirmed with principal component analysis (PCA). Results were compared with widely used software including geNorm, BestKeeper, and NormFinder. Results indicated that GAPDH was the most stable HKG and RPL32 was ranked as the second best. 18S and 28S were found to be the least stable. The proposed model, PCA, geNorm, and BestKeeper were in agreement in detecting the most stable and the least stable HKGs in the equine endometrium during the estrous cycle and early pregnancy.