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Simple Summary: In the abdominal testes of most mammals, with the exception of elephants and a few animal species, spermatogenesis is characterised by an arrest of spermatogenesis due to heat stress. However, the mechanisms underlying normal spermatogenesis in the elephant abdominal testis remain unknown. We hypothesised that comparing the expression of heat stress tolerance molecules of elephant testes with that of other animals and investigating the mechanism of spermatogenesis in elephants might provide clues to overcome the causes of heat stress-induced spermatogenesis dysfunction. The results showed that the elephant's body cells, other than the testes, have a similar mechanism to those of other mammals but that the spermatogenic cells have a different mechanism, with different molecular immunoexpression under heat stress in the abdomen. These findings suggest that these molecules and their relationships may be important in future investigations of the mechanisms that overcome heat stress to maintain spermatogenesis in elephants. The abdominal testes of Asian elephants show normal spermatogenesis. Heat shock in cryptorchid testes elevates heat shock factor (HSF) expression, leading to germ cell apoptosis, while increased heat shock proteins (HSPs) levels provide protection. To investigate how heat shock affects elephant spermatogenic cells, focusing on heat shock-related molecules and the cell death mechanism, immunohistochemistry and TUNEL staining were employed to assess the immunoexpression of several heat shock-related molecules and the status of apoptosis in elephant fibroblasts (EF) induced by heat shock stimulus. Additionally, the immunoexpression of heat shock-related molecules and cell proliferation status in the elephant spermatogenic cells. Our finding indicated that heat shock-induced HSF1 immunoexpression in EF leads to apoptosis mediated by T-cell death-associated gene 51 (TDAG51) while also upregulating HSP70 to protect damaged cells. In elephant spermatogenic cells, immunostaining revealed a predominance of proliferating cell nuclear antigen (PCNA)-positive cells with minimal TDAG51- and TUNEL-positive cells, suggesting active proliferation and apoptosis suppression during normal spermatogenesis in the abdominal testis. Interestingly, spermatogonia co-immunoexpressed HSF1 and HSP90, potentially reducing apoptosis through protective mechanisms different from those observed in other mammals. Spermatogenic cells did not show immunolocalisation of HSP70, and hence, it may not contribute to protecting the spermatogonia from heat shock because the transcriptional activity of HSF1 is suppressed by HSP90A binding. This study provides insight into the specific heat shock response and defence mechanisms in elephant spermatogenic cells and may contribute to our understanding of species-specific adaptation to environmental stresses of the testis. [ABSTRACT FROM AUTHOR]

Saltlicks are fundamental resources for wild Asian elephants (Elephas maximus). This study aimed to assess the nutrients found in natural saltlicks (NSs) and artificial saltlicks (ASs), as well as general soils (GS) in the natural forest of Salakphra Wildlife Sanctuary (SWS) and a restoration area of Kui Buri National Park (KNP), a which is a forest in Western Thailand. We monitored 33 NSs, 35 ASs, and 20 GSs used by wild Asian elephants. In both areas, the K, Mg, Fe, and Cu in NSs were significantly higher than in ASs. The Ca and Zn in NSs of KNP were lower than the ASs of SWS. The salinity of ASs was the highest, making it significantly higher than that of the NSs in both areas. The ASs can supplement Na, thereby increasing salinity in both areas. The Ca, K, Mg, Fe, and Cu in NSs were significantly higher than in ASs, making them a primary target for elephants. These findings have consequences for conserving elephants and other large herbivores by supplementing essential macro- and micro-nutrients in ASs. [ABSTRACT FROM AUTHOR]

Abstract Background Artificial insemination (AI) using frozen-thawed semen is well established and routinely used for breeding in various mammalian species. However, there is no report of the birth of elephant calves following AI with frozen-thawed semen. The objective of the present study was to investigate the fertilizing ability of chilled and frozen-thawed semen in the Asian elephant following artificial insemination (AI). Methods Semen samples were collected by from 8 bulls (age range, 12-to 42-years) by manual stimulation. Semen with high quality were either cooled to 4°C or frozen in liquid nitrogen (-196°C) before being used for AI. Blood samples collected from ten elephant females (age range, 12-to 52-years) were assessed for estrus cycle and elephants with normal cycling were used for AI. Artificial insemination series were conducted during 2003 to 2008; 55 and 2 AI trials were conducted using frozen-thawed and chilled semen, respectively. Pregnancy was detected using transrectal ultrasonography and serum progestagen measurement. Results One female (Khod) inseminated with chilled semen became pregnant and gave birth in 2007. The gestation length was 663 days and the sex of the elephant calf was male. One female (Sao) inseminated with frozen-thawed semen showed signs of pregnancy by increasing progestagen levels and a fetus was observed for 5 months by transrectal ultrasonography. Conclusion This is the first report showing pregnancy following AI with frozen-thawed semen in the Asian elephant. Successful AI in the Asian elephant using either chilled or frozen-thawed semen is a stepping stone towards applying this technology for genetic improvement of the elephant population.

Abstract Background One of the major obstacles in using artificial insemination to manage genetics of elephant population in captivity is the large variations in semen quality among ejaculates within the same and among individuals. The objectives of this study were to determine the influences of (1) age (2) seasonality (3) and circulating testosterone (SrTest), triiodothyronine (SrT3) and tetraiodothyronine (SrT4), as well as seminal (4) testosterone (SpTest), zinc (SpZn) and protein (SpTP) on semen quality in the Asian elephant Methods Analyses, including motility, viability and morphology were performed in semen samples collected twice monthly from 13 elephant bulls (age range, 10-to 72-years) by manual stimulation between July 2004 and June 2005. Serum samples obtained monthly were assessed for SrTest, SrT3, SrT4, and seminal plasma samples were evaluated for, SpTest, SpZn and SpTP. Results The highest semen quality was observed at age 23 to 43 years. Percentages of progressive motility and viable sperm were lowest at age 51 to 70 years (P < 0.05); on the other hand, sperm concentration was lowest at age 10 to 19 years (P < 0.05). Percentage of sperm with normal morphology was highest at age 23 to 43 years. The levels of SrT3, SrTest, SpTest and SpZn were lowest at age 51 to 70 years, whereas SrT4 was lowest at age 23 to 43 years. Seasonality significantly affected semen characteristics in which percentage of viable sperm and cell concentration were highest during rainy season and lowest during summer months (P < 0.05). However, percentage of sperm with normal morphology was highest in summer and lowest in rainy season (P < 0.05). Seasonality significantly influenced SrTest with elevated concentrations observed in rainy season and winter (P < 0.05). Conclusion This study indicates that age and seasonality had influence on semen characteristics in the Asian elephant. The knowledge obtained in this study will improve our understanding of the reproductive biology of this species.

A captive female Asian elephant (Elephas maximus), approximately 30 yr old, was struck by lightning and displayed obtundation, dysphagia, drooling, and muscle weakness. A physical examination, hematologic test, blood gas analysis, and electrocardiogram were performed. Treatments included the administration of mannitol, dexamethasone, antibiotics, analgesics, vitamins, and intravenous fluid. Conventional therapy resulted in improvement but not complete resolution of clinical signs. Therefore, acupuncture was integrated into the treatment protocol. Electroacupuncture and laser acupuncture were administered every other day; aqua acupuncture was administered once a week. No complications were observed during acupuncture treatment. Acupuncture, a form of traditional Chinese veterinary medicine (TCVM), contributed to the resolution of clinical signs in this case and may be an effective adjunctive treatment for other neurologic disorders in elephants. [ABSTRACT FROM AUTHOR]

Contents: The aim of this study was to evaluate home‐made and commercial extenders for the cryopreservation of Rusa deer semen. After collection by electroejaculation, six ejaculates were diluted and frozen in TES‐based, Tris‐based and Triladyl® extenders. Subjective motility, viability, morphology, acrosome integrity and membrane functionality were assessed post‐thawing and after 1‐hr incubation at 37°C (Thermal stress test). Total and progressive motility, and kinematic parameters were also assessed through CASA system. Post‐thawing sperm progressive motility (PM), velocity according to the straight path (VSL) and linearity (LIN) showed significant differences, and higher values were detected for spermatozoa diluted with Triladyl® and TES (p < 0.05) as compared with Tris (PM of Triladyl® 14.7% vs. 3.2% TES and 2.5% Tris; VSL 56 for Triladyl®, 59.2 for TES and 41.7 for Tris; LIN 45.6 for Triladyl®, 52 for TES and 36.5 for Tris). Triladyl® and TES extender led to better post‐thawing sperm parameters, but these preliminary results need to be verified through artificial insemination trials. [ABSTRACT FROM AUTHOR]

The Asian tapir (Tapirus indicus) has been classified as Endangered on the IUCN Red List of Threatened Species (2008). Genetic diversity data provide important information for the management of captive breeding and conservation of this species. We analyzed mitochondrial control region (CR) sequences from 37 captive Asian tapirs in Thailand. Multiple alignments of the full-length CR sequences sized 1268 bp comprised three domains as described in other mammal species. Analysis of 16 parsimony-informative variable sites revealed 11 haplotypes. Furthermore, the phylogenetic analysis using median-joining network clearly showed three clades correlated with our earlier cytochromebgene study in this endangered species. The repetitive motif is located between first and second conserved sequence blocks, similar to the Brazilian tapir. The highest polymorphic site was located in the extended termination associated sequences domain. The results could be applied for future genetic management based in captivity and wild that shows stable populations. [ABSTRACT FROM PUBLISHER]

A 45-year-old female Asian elephant (Elephas maximus) was presented with urinary incontinence and solid masses at the perineal area. Vaginoscopy revealed stricture of the vaginal vestibule. Trans skin and rectal ultrasonography demonstrated presence of hyperechogenic uroliths. Episiotomy and urethrotomy were performed after standing sedation with xylazine and perineal infiltration with lidocaine. Total of 8 kg urethral calculi were retrieved manually. Calcium carbonate was identified as the main component of the stones. Due to acute urinary obstruction after the operation, an episiotomy was left open allowing urine to pass through the incision. Following urine scalding, necrotic tissues were detected along the tracts below the incision wound through umbilicus. Daily wound cleaning and antibiotics treatment for one month were performed. The episiotomy wound was left to heal by secondary intention and led to a 2 cm stricture. The healing process completed in 5 months post-operatively. The elephant can urinate via normal tract in 8 months after surgery. [ABSTRACT FROM AUTHOR]

The abdominal testes of Asian elephants show normal spermatogenesis. Heat shock in cryptorchid testes elevates heat shock factor (HSF) expression, leading to germ cell apoptosis, while increased heat shock proteins (HSPs) levels provide protection. To investigate how heat shock affects elephant spermatogenic cells, focusing on heat shock-related molecules and the cell death mechanism, immunohistochemistry and TUNEL staining were employed to assess the immunoexpression of several heat shock-related molecules and the status of apoptosis in elephant fibroblasts (EF) induced by heat shock stimulus. Additionally, the immunoexpression of heat shock-related molecules and cell proliferation status in the elephant spermatogenic cells. Our finding indicated that heat shock-induced HSF1 immunoexpression in EF leads to apoptosis mediated by T-cell death-associated gene 51 (TDAG51) while also upregulating HSP70 to protect damaged cells. In elephant spermatogenic cells, immunostaining revealed a predominance of proliferating cell nuclear antigen (PCNA)-positive cells with minimal TDAG51- and TUNEL-positive cells, suggesting active proliferation and apoptosis suppression during normal spermatogenesis in the abdominal testis. Interestingly, spermatogonia co-immunoexpressed HSF1 and HSP90, potentially reducing apoptosis through protective mechanisms different from those observed in other mammals. Spermatogenic cells did not show immunolocalisation of HSP70, and hence, it may not contribute to protecting the spermatogonia from heat shock because the transcriptional activity of HSF1 is suppressed by HSP90A binding. This study provides insight into the specific heat shock response and defence mechanisms in elephant spermatogenic cells and may contribute to our understanding of species-specific adaptation to environmental stresses of the testis.

Background and Aim: Brucellosis is considered as an important zoonotic disease caused by various strains of Brucella in numerous host species. Although brucellosis has been reported in almost animal species, the relevance of brucellosis infection and diagnostic technique in Asian elephant ( Elephas maximus ) has been limited. The present serological investigation aimed to investigate the antibody response to Brucella abortus in captive Asian elephants in North Thailand. Moreover, further serological survey was also conducted to detect the antibody response to Brucella canis in stray dogs cohabiting the same area as the elephant herd., Materials and Methods: Serum samples were collected from 40 captive Asian elephants and submitted for serological analysis based on B. abortus antigen using Rose Bengal plate test (RBPT) in combination with ethylenediaminetetraacetic acid-tube agglutination test (EDTA-TAT) as a supplementary test and by commercial indirect enzyme-linked immunosorbent assay (iELISA). In addition, serum samples were also obtained from 16 stray dogs that live nearby the elephant-raising area and were tested using commercial Dot-ELISA based on B. canis antigen., Results: Serological analysis in captive Asian elephants showed 100% seronegative (40/40) from all serological tests response to B. abortus . For stray dogs, 12.5% (2/16) had a low positive reaction response to B. canis ., Conclusion: The serological survey for brucellosis in Asian elephant was adapted and applied using RBPT, EDTA-TAT, and iELISA in the present study. For future evaluation, we recommended the use of a combination of serological tests with validation together with comparing by direct detection such as bacterial isolation to provide an appropriate brucellosis surveillance program in Asian elephants. In addition, the surveillance of stray dogs or multispecies habitation should be kept into considerations., (Copyright: © Ngamkala, et al.)

To clarify the reproductive cycle of female Rusa deer (Rusa timorensis), the fecal concentrations of progesterone and 17β-estradiol metabolites were measured. Fecal samples were collected on a weekly basis for one year (between October, 2012 and September, 2013) from five healthy adult hinds in Thailand. At the beginning of the study, three hinds were pregnant. Two hinds delivered one healthy offspring, and one hind delivered a stillborn calf. The mating period of Rusa hinds in Thailand is from November to April. In pregnant hinds, fecal progesterone metabolite concentration was high in late pregnancy and abruptly declined to the baseline around parturition, suggesting that the placenta secretes a large amount of progesterone. Fecal 17β-estradiol metabolite concentration remained elevated around the day of parturition. Both concentrations of fecal progesterone and 17β-estradiol metabolites in non-lactating hinds were significantly higher than those in lactating hinds, indicating that ovarian activity of lactating hinds is suppressed by the suckling stimulus of fawn during lactation. The present study demonstrated that monitoring of fecal steroid hormones is useful method for assessing ovarian function in this species.