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Objective: The skin microbiome plays a role in the pathogenesis of acne vulgaris (AV). Among other species, Malassezia spp. can be found in the pilosebaceous follicle. However, its role on the pathogenesis of AV has not yet been described. The objective of this study was to identify the presence and the distribution of Malassezia spp. in facial AV lesions and also to compare the distribution of Malassezia spp. between inflammatory and non-inflammatory lesions. Methods: One hundred and twenty subjects were allocated into two groups: inflammatory lesions and non-inflammatory lesions. Samples were taken from hair follicles and examined by microscopy using 20% potassium hydroxide and Parker ink and cultured for species identification. Results: There was no difference in the spore load between the two groups (p=0.142). Malassezia spp. were isolated from 64.2% of subjects and consisted of M. dermatis (43%), M. sympodialis (18%), M. slooffiae (16%), M. japonica (5%), M. furfur (5%), M. pachydermatis (3%), and M. restricta (1%). There was a higher Malassezia spp. proportion in non-inflammatory lesions (p=0.019). The predominant species in inflammatory AV lesions was M. dermatis (45,8%), followed by M. sympodialis (17.1%), and M. slooffiae (11.4%). Conclusion: Malassezia spp. were found in facial acne lesions. M. dermatis was the predominant species found in facial AV, followed by M. sympodialis, and M. slooffiae. A higher proportion of Malassezia spp. was found in non-inflammatory lesions.

Spontaneous preterm labor is frequently caused by an inflammatory response in the gestational tissues elicited by either infectious or sterile agents. In sterile preterm labor, the key regulators of inflammation are not identified, but platelet-activating factor (PAF) is implicated as a potential rate-limiting effector agent. Since Toll-like receptor (TLR)-4 can amplify PAF signaling, we evaluated whether TLR4 contributes to inflammation and fetal loss in a mouse model of PAF-induced sterile preterm labor, and whether a small-molecule TLR4 inhibitor, (+)-naltrexone, can mitigate adverse PAF-induced effects. The administration of carbamyl (c)-PAF caused preterm labor and fetal loss in wild-type mice but not in TLR4-deficient mice. Treatment with (+)-naltrexone prevented preterm delivery and alleviated fetal demise in utero elicited after cPAF administered by i.p. or intrauterine routes. Pups born after cPAF and (+)-naltrexone treatment exhibited comparable rates of postnatal survival and growth to carrier-treated controls. (+)-Naltrexone suppressed the cPAF-induced expression of inflammatory cytokine genes Il1b, Il6, and Il10 in the decidua; Il6, Il12b, and Il10 in the myometrium; and Il1b and Il6 in the placenta. These data demonstrate that the TLR4 antagonist (+)-naltrexone inhibits the inflammatory cascade induced by cPAF, preventing preterm birth and perinatal death. The inhibition of TLR4 signaling warrants further investigation as a candidate strategy for fetal protection and delay of preterm birth elicited by sterile stimuli. Refereed/Peer-reviewed

Objective: To evaluate normal spermatozoa morphometry and ultrastructure in estuarine crocodile (Crocodylus porosus). Methods: Four adult male crocodiles aged between 12-15 years, with an average snout to tail length of (3.15±0.01) m were selected for this study. Manipulation of the phallus digitally from the base of the penis was performed on four adult male crocodiles to facilitate the flow of semen to the sulcus. Semen was collected from all individuals for spermatozoa morphometry and ultrastructure study. Morphometry analysis was performed from eosin-nigrosin stained spermatozoa sample. Scanning electron microscopy was conducted to observe the surface ultrastructure of spermatozoa. Results: The morphology of crocodile spermatozoa was made up of acrosome, head, and tail which corresponded to (5.55±1.20) μm, (12.74±1.57) μm, and (70.67±4.40) μm, respectively. The total length of spermatozoa in estuarine crocodile was measured at (88.96±0.52) μm. The most common spermatozoa abnormalities found in the samples were cytoplasmic droplets. Conclusions: Normal morphology, morphometry of spermatozoa in estuarine crocodile which are vermiform shaped-head with a long tail along with questionably high cytoplasmic droplets count are described in this study.

Caseous lymphadenitis (CLA) is a chronic bacterial disease that causes economic losses in small ruminant throughout the world. Corynebacterium pseudotuberculosis is the causative agent of CLA characterized by large abscesses internally/externally, abortions, weight loss, and decreased wool production in sheep and goat farming. There is paucity of information on Corynebacterium pseudotuberculosis and its immunogen mycolic acid responses in male goat during the course of CLA. As a result, this study was designed to investigate the clinical responses and pathological changes of the reproductive tract and its associated lymph nodes in male goats following inoculation with Corynebacterium pseudotuberculosis and its immunogen mycolic acids. A total of 12 healthy crossbred Boer male goats aged 16–20 months were divided into three groups, 1, 2, and 3, of four in each group. Group 1 (control group) was inoculated with 2 ml of sterile phosphate-buffered solution (PBS) via intradermal route, group 2 was inoculated with 2 ml 1 × 109 colony forming unit (cfu) of C. pseudotuberculosis intradermally and group 3 was inoculated with 2 ml of mycolic acid (1 g/ml) intradermally. The bucks were observed and assessed for changes in clinical responses daily for 60 days post challenged. At the end of the experiment, animals were slaughtered and organs of the reproductive tract and lymph nodes were harvested for histopathological investigations. In this study, all experimental bucks survived throughout the study period with some evidence of clinical manifestation such as pyrexia, weight loss, and enlargement or rupture of the lymph nodes. Microscopically, C. pseudotuberculosis and mycolic acid challenged groups showed significant cellular changes in the reproductive system and lymph nodes. Group 2 showed abscessation of the lymph nodes while group 3 did not indicate any abscess formation in the lymph nodes. For mycolic acid extract, the result revealed only mild to moderate clinic-pathological responses in bucks with no abscess formation in the lymph nodes. The result obtained added knowledge in the field of CLA research in small ruminants. In conclusion, this research revealed that C. pseudotuberculosis has a negative effect on reproductive performances of bucks and affects the clinical responses and abscess formation.

Corynebacterium pseudotuberculosis is the causative agent of caseous lymphadenitis, a debilitating chronic disease of sheep and goats. Little is known about the buck's reproductive pathophysiology with respect to inoculation with Corynebacterium pseudotuberculois and its immunogen mycolic acid extract. Therefore, this present study was designed to determine the concentration of testosterone hormone, pro-inflammatory cytokines, and semen quality of the experimental animals. A total of 12 bucks, divided into groups 1, 2, and 3 (Negative control group, Positive control group and Mycolic acid group respectively), were enrolled in this study. Following inoculation, all goats were observed for clinical responses and monitored for 60 days post-challenge and were then sacrificed. Blood samples were collected via the jugular once before inoculation and on a weekly basis post-challenge. Semen samples were collected 2 weeks post-challenge and prior to the sacrifice of the experimental animals. During the post inoculation period of 60 days, the concentration of testosterone hormone for group 2 was increased significantly (p 0.05) in weeks 5, 6, and 9 but decreased in weeks 2 and 7 post inoculation. In group 3, the mean concentration of testosterone was increased significantly (p 0.05) in weeks 5, 6, 7, and 9 post inoculation but decreased in week 2. The concentration of interleukin 6 (IL 6) in treated group 2 did not show any significant difference (p 0.05) but increased significantly (p 0.05) in week 2 post inoculation in group 3. For concentration of interleukin 1β (IL1β) in both treated groups 2 and 3 showed significant difference (p 0.05) in weeks 2 and 3 post inoculation. The tumor necrosis factor-alpha (TNF-α) concentration in both treated groups 2 and 3 did not show any significant difference (p 0.05) as compared to group 1. The concentration of interferon-γ (IFNγ) significantly increased (p 0.05) for group 2 for weeks 2, 3, 4, and 5 where else for group 3 was not in significant difference (p 0.05) compared to group 1. Both group 2 and group 3 showed a reduction in semen qualities as compared to group 1, but the severity was more intense in group 2 if compared to group 3. In conclusion, therefore, the present study concluded that the mycolic acid group revealed significant responses of testosterone hormone concentration, semen quality, and its related pro-inflammatory cytokines in bucks following infection but the severity lesser compared to Corynebacterium pseudotuberculosis group.

Inflammatory activation, a major driver of preterm birth and subsequent neonatal morbidity, is an attractive pharmacological target for new preterm birth therapeutics. Inflammation elicited by intraamniotic infection is causally associated with preterm birth, particularly in infants delivered ≤34 weeks’ gestation. However, sterile triggers of PTB, including placental ischaemic injury, uterine distention, cervical disease, or imbalance in the immune response, also act through inflammatory mediators released in response to tissue damage. Toll-like Receptors (TLRs) are critical upstream gate-keepers controlling the inflammatory activation that precedes preterm delivery, as well as in normal term labour. In particular, TLR4 is implicated for its capacity to sense and integrate a range of disparate infectious and sterile pro-inflammatory triggers, and so acts as a point-ofconvergence through which a range of infectious and sterile agents can activate and accelerate the parturition cascade. Recent studies point to the TLR4 signalling complex as a tractable target for the inhibition of fetal, placental & intraamniotic inflammatory cytokine production. Moreover, studies on mice show that novel small molecule antagonists of TLR4 signalling are highly effective in preventing preterm birth induced by bacterial mimetic LPS, heat-killed E. coli or the TLR4-dependent pro-inflammatory lipid, Platelet Activating Factor (PAF). In this review, we discuss the role of TLR4 in regulating the timing of birth and the potential utility of TLR4 antagonists as novel therapeutics for preterm delivery.

Innumerable Escherichia coli of animal origin are identified, which are of economic significance, likewise, cattle, sheep and goats are the carrier of enterohaemorrhagic E. coli, which are less pathogenic, and can spread to people by way of direct contact and through the contamination of foodstuff or portable drinking water, causing serious illness. The immunization of ruminants has been carried out for ages and is largely acknowledged as the most economical and maintainable process of monitoring E. coli infection in ruminants. Yet, only a limited number of E. coli vaccines are obtainable. Mucosal surfaces are the most important ingress for E. coli and thus mucosal immune responses function as the primary means of fortification. Largely contemporary vaccination processes are done by parenteral administration and merely limited number of E. coli vaccines are inoculated via mucosal itinerary, due to its decreased efficacy. Nevertheless, aiming at maximal mucosal partitions to stimulate defensive immunity at both mucosal compartments and systemic site epitomises a prodigious task. Enormous determinations are involved in order to improve on novel mucosal E. coli vaccines candidate by choosing apposite antigens with potent immunogenicity, manipulating novel mucosal itineraries of inoculation and choosing immune-inducing adjuvants. The target of E. coli mucosal vaccines is to stimulate a comprehensive, effective and defensive immunity by specifically counteracting the antibodies at mucosal linings and by the stimulation of cellular immunity. Furthermore, effective E. coli mucosal vaccine would make vaccination measures stress-free and appropriate for large number of inoculation. On account of contemporary advancement in proteomics, metagenomics, metabolomics and transcriptomics research, a comprehensive appraisal of the immeasurable genes and proteins that were divulged by a bacterium is now in easy reach. Moreover, there exist marvellous prospects in this bourgeoning technologies in comprehending the host bacteria affiliation. Accordingly, the flourishing knowledge could massively guarantee to the progression of immunogenic vaccines against E. coli infections in both humans and animals. This review highlight and expounds on the current prominence of mucosal and systemic immunogenic vaccines for the prevention of E. coli infections in ruminants.

Study in semen of the estuarine crocodiles is limited partly due to the lack of knowledge on the semen collection method and their reproductive biology. Therefore, this study was conducted to determine the possibility of semen collection via digital manipulation of the penis. Four matured male crocodiles kept in the same enclosure were physically restrained for semen collection. The collection was done in January 2015 during crocodiles mating season. Mean (± SEM) seminal volume, mass movement, pH, sperm concentration, general motility, forward movement, rotating and vibrating movements measured were 0.55 ± 0.12 ml, 2.75 ± 0.48, 7.13 ± 0.24, 396.88 ± 43.41 × 106 sperm/ml, 45 ± 17.56%, 37.13 ± 5.12%, 9.56 ± 5.38% and 53.13 ± 1.92% respectively. The most important sperm abnormalities that requires further investigation is the cytoplasmic droplets which was 28.25 ± 2.29%. Sperm motility trend was best preserved in extender made up of 0.01 M PBS with 5% egg yolk. Sperm stored 4 °C in this extender had survived longer than the other two extender which was 11 days. There is a unique trend in the spike of spermatozoa motility post 48 h in chilled condition however the cause of such pattern is still unknown. Semen collection by digital manipulation of the penis was adequate in obtaining semen and the semen characteristics of the estuarine crocodile are described.

The aim of this study was to evaluate the effects of 8% virgin coconut oil (VCO) combined with different percentages of egg yolk in Tris extender on the quality of chilled and frozen-thawed bull semen. A total of 24 ejaculates from four bulls were collected using an electroejaculator. Semen samples were diluted with 8% VCO in Tris extender which contained different concentrations 0% (control), 4%, 8%, 12%, 16% and 20% egg yolk. The diluted semen samples were divided into two fractions: one was chilled and stored at 4°C until evaluation after 24, 72, and 144h; the second fraction was processed by chilling for 3h at 4°C to equilibrate, then packaged in 0.25ml straws and frozen and stored in liquid nitrogen at -196°C until evaluation after 7 and 14 days. Both chilled and frozen semen samples were then thawed at 37°C and assessed for general motility using computer-assisted semen analysis (CASA), viability, acrosome integrity, and morphology (eosin-nigrosin), membrane integrity (hypo-osmotic swelling test) and lipid peroxidation (thiobarbituric acid-reactive substances (TBARS)). The results indicate treatments with 8%, 12%, 16% and 20% egg yolk with 8% VCO had greater sperm quality (P

Aim: The objective of this research is to report parameters for breeding soundness evaluation, semen extension, and cryopreservation in Rusa timorensis. Materials and Methods: Seven healthy stags were chosen for semen collection using an electroejaculator. The collections were performed twice in a breeding season between February and June 2016. Samples were collected between 2 and 3 weeks interval, collected twice for each animal. Semen was evaluated, extended, and cryopreserved using four different extenders; Andromed®, BioXcell®, Triladyl®, and a modified Tris-egg yolk combined with Eurycoma longifolia Jack. Results: R. timorensis semen characteristics according to volume (ml), color, sperm concentration (106/ml), general motility (%), progressive motility (%), and % morphology of normal spermatozoa are 0.86±0.18 ml, thin milky to milky, 1194.2±346.1 106/ml, 82.9±2.8%, 76.1±4.8%, and 83.9±4.8%, respectively. Conclusion: Semen characteristics of R. timorensis collected by electroejaculation is good allowing for cryopreservation and future artificial insemination work. The most suitable extender for Rusa deer semen is Andromed®.

BACKGROUND Acne vulgaris is a pilosebaceous disorder. Bacterial activity and inflammation both influence acne formation. Antibiotics suppress the bacterial activities and elicit anti-inflammatory effects. The overuse of antibiotics may lead to resistance in bacteria. This study was aimed to provide an overview of bacteria that may cause acne and determine their susceptibility to antibiotics.METHODS This was a cross-sectional study sampling from 93 patients with acne in Ciptomangunkusumo Hospital. Comedones were extracted and cultured on Brucella blood agar, under aerobic and anaerobic conditions at 35oC for 24–48 hours. Bacterial identification was performed using Vitek®, and susceptibility test using E-test. Data interpretation was based on the Clinical and Laboratory Standards Institute 2015.RESULTS Staphylococcus epidermidis (50.5%), Propionibacterium acnes (11.0%), and Staphylococcus aureus (7.7%) were identified. Bacteria were not found in 69.2% and 1.1% of samples in anaerobic and aerobic cultures, respectively. P. acnes was susceptible to doxycycline (100%) and minocycline (100%), while 10% was resistant to erythromycin, clindamycin, and tetracycline. S. epidermidis was susceptible to minocycline (100%); but resistant to erythromycin (65.2%), clindamycin (52.2%) and tetracycline (32.6%). The susceptibility of S. epidermidis to doxycycline was 89.1%, which was lower than that of P. acnes (100%). S. aureus was found to be sensitive to minocycline (100%), doxycycline (71.4%), clindamycin (71.4%), and tetracycline (71.4%); but it was resistant to erythromycin (42.9%).CONCLUSIONS Doxycycline and minocycline showed 100% effectiveness for P. acnes. The isolated bacteria were more susceptible to doxycycline compared to tetracycline. The use of clindamycin and erythromycin needs to be limited as most S. epidermidis isolates were resistant to both.

Lead acetate (LA) is a toxic compound that has detrimental effects on the male reproductive system, such as decreased testicular size and function, low androgen hormone concentration, and altered testicular histology. While Nigella sativa (NS) has been shown to possess a handful of therapeutic benefits, few studies have shown its effect on damage caused by lead acetate toxicity in the male reproductive system. In this study, 75 Sprague-Dawley rats were divided into 3 groups of 25 rats, which were further subdivided into 5 subgroups each. Group 1 (negative control) was given distilled water, group 2 (positive control (PC)) administered 10 mg/kg of lead acetate (LA) orally/daily, while groups 3 (T1), 4 (T2), and 5 (T3) were given LA 10 mg/kg and graded concentrations (100, 150, and 200 mg/kg) each of NS. One group each, comprising of 25 rats, was euthanized at days 30, 60, and 90 for collection of blood plasma, epididymis, and organ tissues fixed in 10 % buffered formalin at each time interval. The right caudal epididymis was homogenized and used for the determination of spermiogram. Plasma was used for the determination of testosterone (TS), follicle stimulating hormone (FSH), and luteinizing hormone (LH) and estradiol (EST) using radioimmunoassay kits. There was reduced number of spermatogenic cells and epididymal sperm reserves in the PC group in comparison to the treatment groups. The level of TS was lower (p

The aim of this study was directed towards correlating the body condition score with estrus and pregnancy rate in Nellore cattle. Eightythree cows were randomly selected for the study. Cows were observed for estrus and artificially inseminated. Blood samples were collected twice weekly after estrusduring a whole estrous cycle of 21 days. Cows were scored for body condition based on a 1 to 9 scale. According to body condition score (BCS), cows were subdivided into five groups: BCS 2 (5 cows), BCS 3 (11 cows), BCS 4 (34cows), BCS 5 (26 cows) and BCS 6 (7 cows). Cows with BCS 5 exhibited higher estrus(88.4 %) and pregnancy rate (46.1 %) compared to other cows with BCS of 2, 3, 4 and 6. Progesterone profile was normal in cows with BCS 3, 4, 5 and 6. However, progesterone profile in cows with BCS 2 was delayed and abnormal. In conclusion, higherincidence of estrus and pregnancy rate were observed in the moderate body condition cows (BCS 5). Therefore, management of cows herds should be designed to obtain and maintain cows at a BCS of 5 or greater from pre-calving through rebreeding by adjusting a good feeding program.

Tobacco extract had been proven to have antimicrobial activity against Streptococcus mutans. To maximize the use of tobacco extract on topical use as an antibacterial, it could be formulated into a pharmaceutical preparation. This study aimed to formulate tobacco extract in gel preparation by using Hidroxy Propyl Methyl Cellulose (HPMC) as a gelling agent and to test its activity on S. mutans . The tobacco extract gel was prepared in 3 formulas with variations of HPMC concentration of 1.5% (F1), 2% (F2) and 2.5% (F3). The research began with tobacco extraction, then continued with characteristics evaluation. The extract was formulated in gel form and evaluated for 12 weeks of physical stability. Antibacterial activity was tested using the diffusion method. The evaluation results of tobacco extract gel showed that all formulas were stable during 12 weeks storage. Antimicrobial activity against S.mutans showed inhibitory diameter of F1, F2 and F3, were 9,07 mm, 19,53 mm, and 11,57 mm respectively. The test was continued by determining the relative potential of F2 compared to erythromycin. The test results showed 1.2 x 10 -2 relative potential compare to erythromycin. Based on the results of this study, it can be concluded that HPMC concentration difference did not give significant difference to the physical stability of tobacco gel, with the best antibacterial activity on S. mutans obtained from F2.

Mastitis is the inflammation of the mammary gland due to microbial infiltration causing a reduced mammary function. This study aims at developing a vaccine using Malaysian local isolate of Staphylococcus aureus and evaluating serum amyloid A, Interleukin-10, IgM and IgG responses periodically. Four bacterin concentrations (106, 107, 108 and 109 cfu/ml of the local isolate of S. aureus) were adjuvanted with aluminium potassium sulphate. Thirty cows grouped into 4 treatment groups (G-) were vaccinated (2 ml) intramuscularly, with a fifth G-A as control. The mean concentration (MC) of serum amyloid A (SAA) was significantly different (sig-d) (p ˂ 0.05) in G-D at 0 h post vaccination (PV), 3 h PV, 24 h PV, weeks 1, 2, 3 and 4 PV (6-, 15-, 5-, 12-, 11-, 4- and 11-fold increased (FI) respectively). The MC of serum amyloid A was also sig-d in G-E at 0 h PV, weeks 1, 2 and 4 PV (3, 8, 5 and 8 FI respectively). The MC of IL-10 was sig-d in G-D and C at 3 h PV and week 2 PV (5 and 2 FI respectively). The IgM MC was sig-d in G-B and C at 3 h PV (5 and 6 FI respectively), at 24 h PV (5 and 9 FI respectively), at week 3 PV(2 and 2 FI respectively) and week 4 PV (3 and 4 FI respectively). The MC of IgG was sig-d in G-E at 0 h, 3 h and week 3 PV(5, 6 and 2 FI respectively) and in G-D at weeks 1–4 (3, 3, 3 and 5 FI respectively). In conclusion, elevated levels of SAA, IgG and IL-10 in G-D(108) informed our choice of best dosage which can be used to evoke immunity in cows.

Mastitis is an inflammatory condition of the udder that occurs as a result of the release of leucocytes into the udder in a response to bacterial invasion. The major causes of mastitis are an array of gram positive and negative bacteria, however, algae, virus, fungi, mechanical or thermal injury to the gland have also been identified as possible causes. Mastitis vaccines are yet to be developed using Malaysian local isolate of bacteria. The objective of the present experimental trial was to develop a monovalent vaccine against mastitis using S. aureus of Malaysian isolate and to evaluate the clinical responses such as temperature, respiratory rates and heart rates in vaccinated cows. S. aureus is a major causative bacteria in clinical and subclinical types of mastitis in cows. Four concentrations of the bacterin (106, 107, 108 and 109 cfu/ml of the local isolate of S. aureus) were prepared using Aluminium potassium sulfate adjuvant. Thirty cows were grouped into four treatment groups (B, C, D and E) with a fifth group as control (A). These groups were vaccinated intramuscularly(IM) with the prepared monovalent vaccine and its influence on the vital signs were intermittently measured. The mean of rectal temperature was significantly different (p˂ 0.05) at 0hr Post Vaccination [1]" in groups D and E (39.5 ± 0.15 °C and 39.4 ± 0.15 °C respectively) and at 3 h PV in groups C, D and E (39.8 ± 0.14 °C, 39.9 ± 0.14 °C and 40.3 ± 0.14 °C respectively) compared to the control group. This indicated a sharp increased rectal temperatures between 0hr and 3 h PV in groups C, D and E which later declined at 24 h PV. The mean of rectal temperature of group E was significantly different (p˂ 0.05) at weeks 1 and 2 PV (39.87 ± 0.19 °C and 39.80 ± 0.18 °C respectively) compared to the control group. The mean of heart rate was significantly different (p˂ 0.05) at week 1 PV in groups D and E (83.0 ± 3.8 beats/minute and 80.0 ± 3.8 °C respectively) compared to control. A trending decrease was however observed in heart rates of group E from weeks through 4 PV and in group D from weeks 1 through 3 PV. The mean of respiratory rates was significantly different (p˂ 0.05) at week 3 PV in group B and D (31.0 ± 1.2 breaths/minute and 28.0 ± 1.2 breaths/minute) compared to control. In conclusion, this study highlights responses of these vital signs due to vaccination against S. aureus causing mastitis in cows. To the best of our knowledge the findings of this study adds value to the shallow literature on vital signs alterations in cows vaccinated against mastitis as elevated levels of temperature and heart rates of group D and E indicated obvious response.

Gonado-hypophyseal lesions and reproductive hormonal changes in Brucella melitensis-infected mice and its lipopolysaccharide (LPS) were investigated. Forty-eight mice (24 males and 24 females) were used as infection models in this study, and they were divided into three equal separate groups of eight mice based on their sex: M1, M2, and M3 and F1, F2, and F3 (M = Male, F = Female). Groups M1 and F1 were orally inoculated with sterile phosphate-buffered saline (PBS) and served as the negative control. Groups M2 and F2 were orally inoculated with 0.4 ml of 109 colony-forming units (cfu) of B. melitensis, while groups M3 and F3 were orally inoculated with 0.4 ml of LPSs from 109 cfu of B. melitensis. All the groups were observed for 10 days for changes in clinical signs or signs of mortality and then euthanized. Whole blood was collected via cardiac puncture, while tissues were fixed in 10 % formalin for histopathological processing. Histopathological lesions in the male and female reproductive tracts ranged from mild to moderate degenerative, inflammatory, and necrotic lesions. These changes were moderate in M2 than in M3 in males and in F3 than in F2 groups in females. Lesions in the pituitary gland ranged from mild to moderate inflammation and necrosis with astrogliosis. F2 and M2 groups had more lesion distribution in the pituitary gland than F3 and M3 groups. There was a 13 % decline in progesterone concentration in F2 and a 44 % decline in F3. Estradiol level declined by 33 % in F2 and slightly increased by 4.2 % in F3. Progesterone-estrogen ratio (P/E) increased by 31 % in F2 and declined by 46 % in F3. Testosterone concentration increased by 74 % in M2 and decreased by 56 % in M3. The study showed that oral administration of B. melitensis affects the pituitary gland and may alter the hypothalamic-pituitary-gonadal axis.

Caseous lymphadenitis (CLA) is a chronic disease caused by Corynebacterium pseudotuberculosis. However, there is a paucity of data about the C. pseudotuberculosis exotoxin, phospholipase D (PLD) response during the course of CLA. Therefore, this study was conducted to observe the clinical signs and the cellular changes after an experimental infection of the C. pseudotuberculosis and phospholipase D challenge. Twenty six crossbred Boer goats aged 12-14 months were divided into 3 groups; the first group n=6 was inoculated with 1ml sterile PBS s.c. as the control. The second group n=10 was inoculated with live C. pseudotuberculosis 1×109 cfu s.c. The third group n=10 was i.v. inoculated with PLD 1mL/20 kg, BW. Both the C. pseudotuberculosis and the PLD treated groups showed a significant increase (p

Trauma is a common problem in Cownose Ray during mating season in both wild and captive rays. Enucleation is indicated when there is an ocular trauma. A 5-year-old female Cownose Ray (Rhinoptera bonasus) from Aquaria of Kuala Lumpur Convention Centre (KLCC) was presented to University Veterinary Hospital (UVH), Universiti Putra Malaysia, with a complaint of protruding left eye, which resulted from crushing into artificial coral during mating season. There were a hyphema in the traumatic left eye, periorbital tissue tear, exposed left eye socket, and multiple abrasions on both pectoral fins. The Cownose was anaesthetized and maintained with isoeugenol and on-field emergency enucleation of the left eye was performed. It was managed medically with postoperative enrofloxacin, tobramycin ointment, and povidone iodine. No suture breakdown and secondary infection were observed at day 7 after enucleation during revisit. At day 24 after enucleation, the Cownose responded well to treatment with excellent healing progression and no surgical complication was observed.

This study was carried out to determine different types of luteal sub-function in relation to pregnancy rate in Nelore cattle. Fifty seven cows were used. Blood samples were collected twice per week for determining progesterone concentration within estrous cycle. Seven progesterone profiles were found normal progesterone profiles, delayed progesterone profiles, insufficient progesterone profiles, short luteal phase, long luteal phase, basal progesterone profile, and non-response to prostaglandin F2 treatment. Normal progesterone profiles (52.8 %) were higher than abnormal progesterone profiles. Therefore, pregnancy rate was significant higher in normal progesterone profile (45.6 %) compare to other abnormal progesterone profiles. In conclusion, Different progesterone profiles were observed in individual animal after AI. In addition to, it was observed that the post insemination progesterone profile has an effect on the pregnancy rates in Nelore cattle. Keywords: progesterone profiles, pregnancy rate, artificial insemination, Nelore cattle

The study was conducted to evaluate the effects of α-linolenic acid (ALA) on frozen-thawed quality and fatty acid composition of bull sperm. For that, twenty-four ejaculates obtained from three bulls were diluted in a Tris extender containing 0 (control), 3, 5, 10 and 15 ng/ml of ALA. Extended semen was incubated at 37°C for 15 min, to allow absorption of ALA by sperm cell membrane. The sample was chilled for 2 h, packed into 0.25-ml straws and frozen in liquid nitrogen for 24 h. Subsequently, straws were thawed and evaluated for total sperm motility (computer-assisted semen analysis), membrane functional integrity (hypo-osmotic swelling test), viability (eosin-nigrosin), fatty acid composition (gas chromatography) and lipid peroxidation (thiobarbituric acid-reactive substances (TBARS)). A higher (p < 0.05) percentage of total sperm motility was observed in ALA groups 5 ng/ml (47.74 ± 07) and 10 ng/ml (44.90 ± 0.7) in comparison with control (34.53 ± 3.0), 3 ng/ml (34.40 ± 2.6) and 15 ng/ml (34.60 ± 2.9). Still, the 5 ng/ml ALA group presented a higher (p < 0.05) percentage of viable sperms (74.13 ± 0.8) and sperms with intact membrane (74.46 ± 09) than all other experimental groups. ALA concentration and lipid peroxidation in post-thawed sperm was higher in all treated groups when compared to the control group. As such, the addition of 5 ng/ml of ALA to Tris extender improved quality of frozen-thawed bull spermatozoa.

This study provides standard information on the attributes of sperm and describes the surface structure of normal and abnormal spermatozoa of Rusa timorensis. Two fertile stags were used as the source of semen collected during the first breeding season commencing from April 5 to July 2, 2012. Another five stags were used as the source of semen collected during the second breeding season commencing from April 1 to June 27, 2013. Semen samples were collected from the stags using an electro-ejaculator. The ejaculate was processed and samples prepared for light and scanning electron microscopy (SEM) according to standard methods. No significant difference (P > 0.05) was found between sperm attributes in comparison between different stags and different months of the fertile seasons. The results of this study have also demonstrated that there are no differences in size, shape and surface structure between spermatozoa of the different stags and different months of the fertile seasons. Sperm attributes (volume, pH, sperm concentration, general motility, progressive motility and viability) were 2.2 ± 0.29 ml, 7.2 ± 0.17, 886.3 ± 39.7 × 106 spermatozoa/ml, 78.7 ± 2.01%, 80.8 ± 1.85% and 83.2 ± 0.85%, respectively. Morphological analysis showed low percentage of abnormal spermatozoa 13.9 ± 2.88%. Scanning electron microscopy revealed spermatozoa which consisted of a flat paddle-shaped head, short neck and a tail, which was subdivided into midpiece, principal piece and endpiece. The average spermatozoon was 66.2 ± 0.69 μm in total length. The flat paddle-shaped head was 7.8 ± 0.28 μm long, 4.2 ± 0.15 μm at its widest width, 2.4 ± 0.18 μm basal width and 0.7 ± 0.02 μm thick. As for the tail, the midpiece length was 13.2 ± 0.14 μm, 0.6 ± 0.04 μm in diameter; the principal piece was 42.6 ± 0.04 μm, and 2.8 ± 0.06 μm for the endpiece. Abnormal spermatozoa such as tapered head, microcephalic head, decapitated spermatozoa and bent tails were observed. Results provide standard information useful for development of strategies for semen cryopreservation and assisted reproductive technology in this species.

The aim of this study was to compare the estrus response and pregnancy rate between the indigenous beef cows of Malaysia; Kedah-Kelantan (KK) and the exotic beef cows; Brangus (BR) following progesterone and prostaglandin-based estrus synchronization treatments. A total of 40 KK and 30 BR open cows were selected and each breed group was randomly divided equally into two. Cows in KK1 and BR1 were treated with estradiol benzoate (Cidirol, 1 mg, im) each at the time CIDR® was inserted (Day 0). Cloprostenol (Estrumate, 250 μg,im) was administered at the time of CIDR® removal on Day 9 while 1 mg of Estradiol Benzoate (EB) was injected on Day 10. On the other hand, KK2 and BR2 cows received intramuscular injections of 500 and 250 ug of cloprostenol, on Day 0 and 11, respectively. All cows were then observed for estrus signs and scanned per rectum for ovulation followed by AI upon detection of estrus. Pregnancy status was diagnosed 45 days after AI. Both treatments were effective in inducing observable estrus in all groups with synchrony of ovulation resulting in CL development and pregnancy. In the progesterone4)ased treatment groups, 84.2% of KK1 and 78.8% of BR1 responded. In the prostaglandingsed treatment groups, KK2 responded with the highest proportion (80.0%) compared with BR2 (50.0%). However, there was no significant difference in rate of ovulation (84.2 vs. 64.3%; 70.0 vs. 42.9%) and pregnancy (31.6 vs. 14.3%; 45.0 vs. 21.4%) among all the four experimental groups. The interval to ovulation from the last treatment time varied significantly among all the treatment groups with a higher variation observed in BR, ranging from 48 h when treated with CIDR to 84 h after treatment with PGF2α. These variations could be explained by the difference in ovarian status at the time of treatment. In conclusion, the result of this data showed KK cows had a better rate of ovulation and pregnancy than BR cows in both treatments though not statistically significant. It can therefore be gathered that KK and BR responded effectively to estrus synchronization and produce acceptable pregnancy rates by both progesterone and prostaglandn-based protocols for breeding and genetic improvement.

The aim of this study was to evaluate the effectiveness of different cryodevices (Open Pulled Straw (OPS), Electron Microscopy Grid (EMG) and cryotop for vitrification of immature bovine oocytes. Polar body, MII stage, survivability and subsequent developmental rates were compared. Only oocytes with 4-5 layers of cumulus cells were used. Oocytes were equilibrated in the first vitrification solution (VS1; HS+10% DMSO+10% Ethylene Glycol (EG)) for 30-45 sec and then in the second vitrification solution (VS2; 20% DMSO+20% EG+0.5 M Sucrose) for 25 sec. Within 30 sec they were mounted on one of the cryodevices and directly plunged into Liquid Nitrogen (LN2) for 10 days. Immature oocytes vitrified using cryotop represented higher rate of polar body extrusion and nuclear maturity (p

The aim of this study was to determine the effect of butylated hydroxytoluene (BHT), a lipid-soluble anti-oxidant added in different concentrations to the Tris egg yolk extenders on semen cytological parameters pre freezing and post thawing (motility, morphology, viability, acrosome integrity and membrane integrity) of Boer goat spermatozoa. A total of 40 ejaculates from four Boer goat bucks were collected using an artificial vagina. Ten replicates of the ejaculates were diluted with a Tris egg yolk based extender which contained various concentrations (0.5mM, 1.0mM, 2.0mM and 3.0mM) of butylated hydroxytoluene while one sample was processed without supplementation of antioxidant and served as control. The diluted semen was cooled at 4°C and loaded into the straw and then stored in liquid nitrogen. It was evident that supplementation of BHT produces positive effect in terms of motility, membrane integrity and acrosome integrity in comparison with the control group in cooled and frozen Boer goat semen. Results showed significant differences in motility, membrane integrity, acrosome integrity and viability of cooled and frozen Boer goat spermatozoa at different concentrations. Motility, membrane integrity, acrosome integrity and viability was significantly higher in all treated groups than the control group (P0.05) while there was no significant differences (P0.05) in morphology trait between all group in cooled semen. However, improvement (P0.05) was observed only in terms of the membrane integrity and acrosome integrity compared to the control and other treated groups in frozen semen. In conclusion, BHT can be used in cryopreservation of Boer goat semen in order to reduce the oxidative stress on spermatozoa.

The effectiveness of different cryodevices (open-pulled straw (OPS), electron microscopy grid (EMG), and Cryotop was evaluated for vitrification of immature bovine oocytes. Polar body, metaphase II stage (MII), survivability, and subsequent developmental rates were determined. Only oocytes with four or five layers of cumulus cells were used. Oocytes were equilibrated in two vitrification solutions - 1: 10% DMSO + 10% ethylene glycol (EG) for 30-45sec and 2: 20% DMSO + 20% EG +0.5M sucrose for 25sec -, mounted on one of the cryodevices and directly plunged into liquid nitrogen for 10 days. Immature vitrified oocytes using Cryotop showed the highest rates of polar body extrusion (PB) and nuclear maturity (MII); 41 and 58% respectively. Vitrified oocytes using OPS and EMG showed 26 and 32%; and 35 and 46% of PB and MII rates, respectively. The highest survivability resulted from Cryotop and EMG groups and no significant difference was found between them. Vitrified oocytes using Cryotop had the highest cleavage and blastocyst rates. All of the mean rates for vitrified immature oocytes were significantly lower than that of control group (P

Problem statement: Vitrification is replacing conventional slow freezing to cryopreserve gametes and embryos especially for in vitro production of embryo in domestic animal species. However, the results are still not satisfactory. The aim of this experiment was to study the effect of different equilibration temperatures on in vitro viability of immature bovine oocytes after vitrification. Approach: Oocytes were obtained from slaughterhouse ovaries. Only grade one oocytes were used. Oocytes were equilibrated in three different temperatures: 32, 37, or 41°C. Immature oocytes were equilibrated in VS1 (7.5 Ethylene Glycol (EG) + 7.5% DMSO) for 10-12 min and then exposed to VS2 (15% EG + 15%DMSO + 0.5M sucrose) for one min. Thereafter oocytes were loaded on hand-made Cryotop and directly plunged into liquid nitrogen. After warming, oocytes were examined for viability, maturation, cleavage and blastocyst production. Results: Oocytes that were equilibrated at 37°C had significantly higher (p

The primary objectives of this study were to investigate incidence of abnormal ovarian cyclicity (AOC) and its type in dairy and beef cows with prolonged postpartum period (90 days) and in heifers that fail to conceive. A total of 53 animals were included in the study: 17 Friesian crosses, 16 Braford crosses, eight Brangus crosses, and 12 local Kedah-Kelantan (KKX) crosses. These animals were initially checked for absence of pregnancy via palpation per rectum. Blood samples for progesterone analysis were obtained twice a week for 2 to 3 months following their spontaneous oestrous cycle, and all animals were rechecked for pregnancy at the end of the study. Progesterone analysis indicated that 33.9% of the total animals were having AOC: 18.9% with cessation of ovarian cyclicity, 9.4% with prolonged luteal phases (PLP), and 5.7% short luteal phases. The highest incidence was observed in Brangus crosses (62.5%), followed by Braford crosses (43.8%), and Friesian crosses (35.3%). In contrast, no AOC was observed in the local KKX breeds, and all of them were found to be pregnant at the end of the study. A significant difference (p0.05) in the incidence of AOC and its type was observed between Kedah-Kelantan crosses and the other breeds. Although not significant (p0.05), Friesian crosses showed a higher percentage incidence of AOC than beef cows (40% vs 36.4%), with major types being PLP (26.7%) in dairy and cessation of ovarian cycle (27.3%) in beef cows. Compared with beef heifers, beef cows showed a higher percentage of AOC (36.4% vs 28.6%) where again, cessation of cyclicity was the predominant abnormality. In conclusion, AOC reflected by abnormal endocrine pattern is a possible cause of reduction in fertility for dairy and beef cows beyond 90 days postpartum and heifers that fail to conceive.

An inflammatory response is instrumental in the physiological process of parturition but the upstream signals initiating inflammation are undefined. Because endogenous ligands for Toll-like receptor 4 (TLR4) are released in late gestation, we hypothesized that on-time labor requires TLR4 signaling, to trigger a cytokine and leukocyte response and accelerate the parturition cascade. In pregnant TLR4-deficient (Tlr4−/−) mice, average gestation length was extended by 13 hours and increased perinatal mortality was seen compared with wild-type controls. Quantification of cytokine and uterine activation gene expression showed that late gestation induction of Il1b, Il6, Il12b, and Tnf expression seen in control placenta and fetal membranes was disrupted in Tlr4−/− mice, and accompanied by a transient delay in expression of uterine activation genes, including prostaglandin F receptor, oxytocin receptor, and connexin-43. Leukocyte populations were altered before birth in TLR4-deficient females, with fewer neutrophils and macrophages in the placenta, and fewer dendritic cells and more regulatory T cells in the myometrium. Administration of TLR4 ligand lipopolysaccharide to pregnant wild-type mice induced cytokine expression and fetal loss, whereas Tlr4−/− pregnancies were protected. The small molecule TLR4 antagonist (+)-naloxone increased mean duration of gestation by 16 hours in wild-type mice. Collectively, these data demonstrate that TLR4 is a key upstream regulator of the inflammatory response acting to drive uterine activation and control the timing of labor. Because causal pathways for term and preterm labor converge with TLR4, interventions to manipulate TLR4 signaling may have therapeutic utility for women at risk of preterm labor, or in postterm pregnancy.

The aims of this study were to evaluate the effects of anti-oxidant butylated hydroxytoluene (BHT), when added at different concentrations into lecithin-based Bioxcell(®) (BX) and two egg-yolk-based; Tris (TY) and citrate (CE) semen extenders, on post-thaw bull sperm quality and oxidative stress. A total of 30 ejaculates from three bulls were collected using an electro ejaculator. Ejaculates were extended with one of the BX, TY and CE extenders, which contained different concentrations (0.0 - control, 0.5, 1.0, 1.5, 2.0 and 3.0mM/ml) of BHT. The extended semen samples were chilled to 4 °C, and then frozen slowly to -196 °C in 0.25 ml straws before being stored in liquid nitrogen for 2 weeks. Results showed that supplementation of BHT improved (P

A study was conducted to profile the plasma progesterone (P4) concentrations and establish the length of oestrous cycle in the Rusa timorensis during the breeding season. Five healthy hinds were selected for peripheral blood sampling twice weekly to gauge the P4 levels by radioimmunoassay, at the start of the breeding season indicated by rutting behaviours of sexually active males. The hinds were polyestrous as proven by the cyclic trend of P4 levels. After the presumptive oestrus indicated by the lowest P4 concentrations (0.20±0.09ng/ml), this ovarian hormone was markedly elevated on day 7 of the cycle (0.78±0.20ng/ml), reached its peak (2.61±0.23ng/ml, P

Contents This study was conducted to evaluate the response of Bali bulls (Bos javanicus) to different semen collection methods and their effects on fresh and post-thawed semen quality. The collection methods employed were electro-ejaculation (EE), transrectal massage (RM) and RM followed by EE (RM + EE). A total of 25 untrained Bali bulls (age between 2 and 4 years old) were subjected to the different semen collection methods. Fresh semen samples from all the 25 bulls were evaluated for volume, pH, general motility, live/dead ratio and abnormality using the conventional method. For fresh and frozen samples collected by EE and RM from 10 bulls, computer-assisted semen analysis system was used for precise quantitative measurement of motility, velocity and forward progression. Accucell photometer was used to measure sperm concentration in all samples, regardless fresh and frozen. Semen samples were obtained 100% of the attempts using EE, 84% using RM and 96% using RM + EE. There were no differences among the collection methods for fresh semen quality characteristics, including motility, morphology and viability, but pH and volume were higher for EE than RM and RM + EE. Higher sperm concentration was observed in semen collected by RM than the other two methods. Different age groups (2–3 and >3–4 years old) of the bulls did not show significant differences in volume, pH, sperm concentration, percentages in motility, live/dead ratio and normal sperm morphology. The quality of semen for general and progressive motility, VAP, VSL and VCL and acrosomal integrity after thawing was higher for RM than EE. In conclusion, Bali bulls appeared to respond best to EE and the combination of RM + EE than RM, as a method of semen collection, with a shorter time of stimulation required. Differences in age of the Bali bulls did not affect the semen quality.

The study aims to present baseline data on the reproductive anatomy of a poorly known tropical deer species, Rusa deer ( Rusa timorensis ). The anatomy of female reproductive system is described using seven uniparous hinds, aged between four and eight years. The various reproductive structures were studied via standard descriptive methods. There was a significant difference in the length and width of both right and left ovaries. The left ovary was slightly larger than the right ovary which indicates that it is physiologically more active. The results of the study showed that the anatomy of female reproductive system of R. timorensis was similar to that observed in domestic ruminants except that the uterus did not have an interconual ligament and this implies that the uterine horns are anchored in such a way that sperm deposited into only one uterine horn of the Rusa deer will be transported to the other uterine horn (interconual transport). Unlike the red brocket deer and pampas deer, the cervix of R. timorensis was characterized by six cervical rings projecting into the cervical canal. This feature should be taken into account when designing effective instrumentation and techniques for transcervical passage of semen during artificial insemination in this species. The results from this study have provided baseline data on the reproductive anatomy of this vulnerable species, and the knowledge generated can be useful in the development of appropriate reproductive techniques in order to increase its population in captivity and also enable easy detection of its reproductive anomalies, thus strategies to propagate and conserve the species can be established. Keywords : Anatomy, Female, Reproductive system, Rusa deer, Timorensis

This study compared the effect of oestrus synchronization using controlled-internal-drug-release-device (CIDR) and equine chorionic gonadotropin (eCG) on reproductive efficiency of Dammar ewes in winter. Data of forty-one ewes were divided into two groups; for long CIDR treatment (12 days, LT, n=25) and for short CIDR treatment (6 days, ST, n=16). The LT group was further divided into 3 subgroups to receive eCG either 48 h before CIDR removal (LT1), or at CIDR removal (LT2). Nevertheless, a third subgroup (LT3) received no eCG and served as the control variable. The ST group was also divided into two subgroups to receive eCG either at CIDR removal (ST1) or to receive no eCG (ST2). Five fertile rams naturally mated the ewes. Then in terms of oestrous responses, the pregnancy and lambing rates were 100 percent (100%) in ewes for all treatments. The onset and duration of oestrous was shorter in LTa1 than the remaining subgroups. Litter size in those of LT2 and LT1 subgroups was greater than those of LT3. Additionally, litter size in those of ST1 was greater than in those of ST2 subgroup. This indicated that eCG injection affected litter size. In conclusion, these results indicated that CIDR could successfully be used for either 6 or 12 days prior to ram introduction without having to administer eCG. However, eCG is important to induce multiple births. Keywords: Oestrus synchronization, sheep, mating, lambing, winter

This study was conducted to determine the effect of antioxidants on standard semen parameters, lipid peroxidation and fertility of Boer goat semen after cryopreservation. Ejaculates from four bucks were collected, evaluated and pooled at 37°C. The pooled semen was diluted with Tris citric acid fructose for washing. Semen samples, which were diluted with a Tris-based extender containing the antioxidant ascorbic acid (8.5mg/ml), butylated hydroxytoluene (2mM), cysteine (5mM) and hypotaurine (10mM) and an extender without antioxidant supplementation were cooled to 4°C and frozen in 0.25 straws with programmable freezer and finally stored in liquid nitrogen. Data (10 replicates) were analyzed by one-way analysis of variance. Mean (±SEM) progressive motility was significantly higher in ascorbic acid than other supplement groups and control samples (P0.05). Best values were observed in ascorbic acid followed by BHT, cysteine, and hypotaurine. Antioxidant supplementation in extender showed significant (P0.05) better values than the control group for sperm membrane integrity, acrosome integrity and viability. The ability of antioxidants to reduce the lipid peroxidation (LPO) after freeze thawing was measured by the formation of malondialdehyde (MDA) using the thiobarbituric acid method. Results showed that addition of antioxidants significantly reduced the rate of LPO in comparison to control (P0.05). Ascorbic acid exhibited better values (1.27±0.28), than butylated hydroxytoluene, cysteine and hypotaurine 1.32±0.42, 2.27±0.16 and 2.38±0.17 respectively, which are significantly better than control (3.52±0.54). Higher pregnancy rate was observed with ascorbic acid followed by butylated hydroxtolune, hypotaurine and cysteine. However, differences in the fertility rate were non-significant with hypotaurine, cysteine and control groups.

Domestic buffaloes (Bubalus bubalis) of swamp type (for draught) and river type (for milk) dominate the tropical Asia with slightly more than 169.4 million. They breed throughout the year and produce two calves every 3 years and are fed mainly on straw, crop residues, and mineral supplements. Assisted reproductive technologies such as artificial insemination and embryo transfer have also been introduced and implemented in buffalo production with less success due to difficulty in detecting estrus. Providing good feed and feeding remain a challenge in buffalo husbandry due to limited feed resources. With improved management and proper breeding, the milk production from buffaloes has increased from 1500 to 5000 l per lactation. The river type buffalo currently produces about 73.2 million tonnes of milk annually.

The distribution of lectin bindings in the testis of the smallest ruminant, lesser mouse deer (Tragulus javanicus), was studied using 12 biotinylated lectins specific for d-galactose (peanut agglutinin PNA, Ricinus communis agglutinin RCA I), N-acetyl-d-galactosamine (Dolichos biflorus agglutinin DBA, Vicia villosa agglutinin VVA, Soybean agglutinin SBA), N-acetyl-D-glucosamine and sialic acid (wheat germ agglutinin WGA, s-WGA), D-mannose and d-glucose (Lens culinaris agglutinin LCA, Pisum sativum agglutinin PSA, Concanavalin A Con A), L-fucose (Ulex europaeus agglutinin UEA I), and oligosaccharide (Phaseolus vulgaris agglutinin PHA-E) sugar residues. In Golgi-, cap-, and acrosome-phase spermatids, lectin-bindings were found in the acrosome (PNA, RCA I, VVA, SBA, WGA and s-WGA), and in the cytoplasm (PNA, RCA I, VVA, SBA, WGA, LCA, PSA, Con A and PHA-E). s-WGA binding was confined to the spermatid acrosome, but other lectins were also observed in spermatocytes. In spermatogonia, VVA, WGA, Con A, and PHA-E bindings were observed. Sertoli cells were intensely stained with DBA and Con A, and weakly with PHA-E. In interstitial Leydig cells, RCA I, DBA, VVA, Con A, PSA, LCA, WGA and PHA-E were positive. UEA I was negative in all cell types including spermatogenic cells. Unusual distribution of lectin-bindings noted in the testis of lesser mouse deer included the limited distribution of s-WGA only in the spermatid acrosome, the distribution of DBA in Sertoli cells, Leydig cells and lamina propria, and the absence of UEA I in all type cells. The present results were discussed in comparison with those of other animals and their possible functional implications.

The Sumatran rhinoceros (Dicerorhinus sumatrensis) is the smallest of all the rhino species. It is one of the rarest mammals in the world and is in imminent danger of extinction. A study was carried out on seven wild-caught females, three wild-caught males and one captive born female Sumatran rhinoceros at the Sumatran Rhinoceros Breeding Centre in Sungai Dusun, Selangor, Malaysia, beginning 1990. As a result of the paucity of scientific information on the reproductive biology of the Sumatran rhinoceros, this study was conducted to obtain information, which could assist in the captive breeding of this endangered and near extinct species. The anatomy of the reproductive system was based on two post-mortem specimens and transrectal real-time ultrasonography in six adult females. Genitalia of the Sumatran rhinoceros were similar to those of other species of rhinoceroses. The cervix consisted of several folds, the uterus was bicornuate with a short body and prominent horns and the ovaries were completely covered by the fimbriated end of the fallopian tube. The internal genitalia could be imaged by ultrasonography. The testes were located within a pendulous scrotum. Two lateral projections were located at the base of the penis. A well-defined process glandis was present at the tip of the penis. The accessory sex glands and the testes could be imaged by ultrasonography.

The delayed domestication of Rusa timorensis may be associated with their poor temperament and to date there is no published information on the temperament of the farmed R. timorensis. Understanding of the R. timorensis temperament and selection program for its evaluation in a breeding herd is important not only for R. timorensis farming but also to other types of animal production. We investigated the temperament of R. timorensis (n = 17) raised in the tropics and determined its relationship with stress. A distance of 13.2 m was fixed for the measurement of flight times. Rusa timorensis hinds with rapid speed are considered temperamental. Each R. timorensis hind was earmarked for a crush test score between 1 and 5; 1 represents calm and 5 represents highly agitated R. timorensis. Stress was determined by measuring plasma cortisol using a cortisol RIA kit and live weight gain was determined by weighing the animals weekly. The hinds were aged using their date of birth records. We found a strong negative correlation between flight time, crush score, and plasma cortisol concentration (P < 0.05). Animals with very poor temperament have elevated plasma cortisol level and lower weight gain (P < 0.05). It was concluded that flight time, crush score, and plasma cortisol concentration could be used for selecting R. timorensis hinds based on temperament for the breeding herd. This method is quick and easy to implement on a farm; therefore, it remains the test choice for selecting animals based on temperament for the breeding herd.

Based on previous studies for vitrification of oocytes, it has been shown that short term exposure to DMSO during vitrification could improve the maturation rate and cause not spontaneous parthenogenesis (Isachenko et al., 2006). In addition, it was reported that DMSO in freezing media caused disassembly of microfilaments and chromosomal abnormalities in mouse oocytes (Vincent et al., 1990). On the other hand, DMSO is categorized as a potent glass former and its existence in vitrification solution seems necessary. The aim of this study was to determine the in vitro viability of immature bovine oocytes vitrified by short or long time exposure to DMSO. Materials and Methods: Cumulus oocytes complexes (COCs) with homogenous ooplasm were recovered from slaughterhouse ovaries and used in this study. The vitrification protocol was adapted from Kuwayama et al (2005) with minor modifications. Briefly, oocytes were washed twice in holding solution (HS, Hepes-buffered TCM medium supplemented with 20% fetal calf serum, FCS) and kept there for about 15 min. Group of four COCs were incubated in the first vitrification solution (VS1; 7.5% DMSO and 7.5% EG in HS) for 12 min. Equilibration in VS1 was performed in three steps of increasing concentration. First (F) and second (S) steps contained 1/3 and 2/3 of VS1 diluted in HS, and the third (T) step contained only pure VS1. Based on removal of DMSO from each step, five treatment groups were designed: (G1) control, (G2) VS1, (G3) F w/o DMSO, (G4) F+S w/o DMSO, and (G5) F+S+T w/o DMSO. For G3, G4 and G5, similar concentration of EG was added to replace DMSO in VS1. All treatment groups were equilibrated into the second vitrification solution (VS2; 15% DMSO, 15% EG and 0.5M sucrose in HS) for a further 60 sec. Two experiments were performed: (a) cytotoxicity after only exposure, and (b) in vitro viability after vitrification processes. In cytotoxicity test, immature oocytes were directly transferred to the warming solution (WS). In vitrification experiment, oocytes were instantly loaded on a Cryotop device and submerged into liquid nitrogen (LN2) for storage. The time of exposure from VS2 to LN2 was not longer than 90 s. Vitrified samples were maintained in LN2 for at least 10 days. Immediately after removing the Cryotop from LN2, thin strip of Cryotop was submerged in 3 ml HS plus 1M sucrose (WS; 39°C) and smoothly tried to detach oocytes from Cryotop device. Immature oocytes were left in WS for one minute and then transferred to HS plus 0.5M and 0.15M sucrose solution for 3 and 5 min, respectively. Finally, the immature oocytes were washed twice in HS for 5 min each and processed for in vitro maturation. Significant differences among treatments used in the experiment were revealed by one-way analysis of variance and followed by Duncan's multiple range test for mean comparisons (P < 0.05) using SAS software (ver. 9.1).

Background: Digestive tract infection with clinical manifestation of diarrhea is an infectious disease that has the highest morbidity and mortality rate, especially in developing countries. Diarrhea causes mortality mostly in infants under one year old. Improvement in management is done continuously, but advances in diagnosis and therapy cannot be reached by developing countries. One of the etiological agents causing infection of digestive tract is bacteria. Therefore, knowledge of bacteria that cause gastrointestinal infection and their resistance patterns may support the management of this disease. The aim of this study was to examine microbes that were isolated from the digestive tract and their resistance patterns against antibiotics.Methods: Samples (stool, rectal/anal swab) were collected from the Clinical Microbiology Laboratory, FKUI during 2005-2008. Isolation, identifi cation and sensitivity test were conducted according to standard laboratory procedures. Interpretation of sensitivity test was done according to NCCLS/CLSI guidance. Data was analyzed using WHOnet version 5.3.Results: We found 28 isolates of pathogenic Escherichia coli, 1 isolate of S. paratyphi A and 4 isolates of yeasts. Pathogenic Escherichia coli were still sensitive against some antibiotics, but the sensitivity was reduced against amoxicillin, sulbenicillin, ticarcillin and trimethoprim/sulfamethoxazole.Conclusion: The most predominant gastrointestinal tract infection causing microbes was pathogenic Escherichia coli. These bacteria showed decrease sensitivity against some antibiotics commonly used to treat patients with gastrointestinal tract infection. (Med J Indones 2011; 20:105-8)Keywords: antibiotics, Escherichia coli, gastrointestinal infection

The spread of drug resistant microbes is a global public health challenge which impairs the efficacy of antimicrobial agents and causes substantial increase in morbidity and mortality rates, including healthcare-associated costs. Monitoring of antimicrobial drug resistance from documented microbial epidemiology & resistance rate is useful in preventing the emergence of resistance. This study reports on the pattern of bacterial resistance against ceftriaxone in the past 4 years. The data were obtained from specimens examined in the Clinical Microbiology Laboratory, Department of Microbiology Faculty of Medicine, University of Indonesia from 2002 to 2005. Microbial species were determined from culture and identification tests. Disc diffusion method was used for sensitivity testing of ceftriaxone to 14 Gram-negative and 7 Gram-positive bacteria. Although resistance rates were increased from 2002 to 2005, resistance rates of ceftriaxone were found to be less than 50%. Low resistance rates (< 3%) were observed for Salmonella typhi, Salmonella paratyphi A, Shigella flexneri, Serratia marcescens, and Streptococcus pneumoniae. These results could be useful in developing guidelines on the use of ceftriaxone in Indonesia. (Med J Indones 2007; 16:3-6) Keywords: Microbial drug resistance, disc diffusion method, Gram-positive, Gram-negative

Aim: The objective of this study was to evaluate the effects of different concentrations of soybean lecithin (SL) and virgin coconut oil (VCO) in Tris-based extender on chilled and frozen-thawed bull semen quality parameters. Materials and Methods: A total of 24 ejaculates were collected from four bulls via an electroejaculator. Semen samples were diluted with 2% VCO in Tris-based extender which consists of various concentrations of SL (1, 1.25, 1.5, and 1.75%). A 20% egg yolk in Tris used as a positive control (C+). The diluted semen samples were divided into two fractions; one for chilling which were stored at 4°C for 24, 72, and 144 h before evaluated for semen quality parameters. The second fraction used for freezing was chilled for 3 h at 4°C, packed into 0.25 mL straws and then cryopreserved in liquid nitrogen. The samples were then evaluated after 7 and 14 days. Chilled and frozen semen samples were thawed at 37°C and assessed for general motility using computer-assisted semen analysis, viability, acrosome integrity and morphology (eosin-nigrosin stain), membrane integrity, and lipid peroxidation using thiobarbituric acid reaction test. Results: The results showed that all the quality parameters assessed were significantly (p