Seroprevalence and contributing factors of transboundary animal diseases in sheep and goats: a study in Peninsular Malaysia.

Diseases caused by small ruminant lentiviruses, Mycobacterium avium ssp. paratuberculosis (MAP), Schmallenberg virus, and peste des petits ruminants virus (PPR) is globally recognised as serious threats to the ruminant industry due to their potential to spread rapidly across boundaries. Despite their global distribution and negative impacts on ruminant production, there is a gap in knowledge of the current trends in their epidemiology among sheep and goat populations in Peninsular Malaysia. This study was therefore designed to fill the gap of knowledge concerning the seroprevalence and contributing factors of CAEV, paratuberculosis, SBV, and PPRV among small ruminants from selected flocks in Selangor, Negeri Sembilan, and Pahang states in Peninsular Malaysia. A cross-sectional study design was used to collect animal data and blood samples for serological assays simultaneously. The ID Screen (ID.VET, France) indirect ELISA screening tests were used to detect serum antibodies directed against CAEV/MVV (VISNAS Ver 0922), paratuberculosis (PARAS Ver 0516), SBV (SBVC Ver 1114) and PPRV (PPRC Ver 0821). There was 45.4% (95% CI = 40.74-50.74), 6.8% (95% CI = 4.66-9.69), 27.8% (95% CI = 23.35-32.77), and 2.6% (95% CI = 1.11-0.51) true seroprevalence for CAEV, paratuberculosis, SBV, and PPR, respectively. Geographical location and species were the risk factors for CAEV and paratuberculosis, while the management system and age of small ruminants were the risk factors for SBV. The present study is the first to document a large-scale seroprevalence of MAP and PPR infection among sheep and goat flocks in Peninsular Malaysia. The presence of PPRV and MAP antibodies among small ruminant flocks is signalling current or previous exposure to the pathogens or cross reactions with similar antigens. This finding further suggests the potential for future outbreaks of these devastating diseases among sheep and goats in Malaysia. The high seroprevalence of CAEV and SBV among small ruminants indicates high levels of exposure to the viruses in the environment, which is a potential threat to production.

Pro-inflammatory cytokines and reproductive hormone responses in bucks post-challenge with Mannheimia haemolytica A2 and its outer membrane protein.

The lipopolysaccharide (LPS) endotoxin and outer membrane protein (OMP) are among the virulence factors of Gram-negative bacteria responsible for inducing pathogenicity in the infected host. OMP and LPS occur on the outer membrane of M. haemolytica A2, the primary aetiological agent of pneumonic mannheimiosis in small ruminants. While the LPS is known to mediate Gram-negative bacterial infection by activating downstream inflammatory pathways, the potential role of OMP during inflammatory responses remained unclear. Hence, this study determined the effect of the OMP of M. haemolytica A2 on the serum concentration of pro-inflammatory cytokines and the male reproductive hormones (testosterone and Luteinizing Hormone). We randomly assigned twelve bucks to three groups (n = 4 bucks each): Group 1 was challenged with 2 mL PBS buffer (pH 7.0) intranasally; Group 2 received 2 mL of 1.2 X 109 CFU/mL whole M. haemolytica A2 intranasally; and Group 3 received 2 mL of OMP extract obtained from 1.2 X 109 CFU/mL M. haemolytica A2 intramuscularly. Serum samples collected at pre-determined intervals were used for the quantitative determination of the pro-inflammatory cytokines (IL-1ß, IL-6, and TNFα) and reproductive hormones (testosterone and LH) using commercial sandwich enzyme-linked immunosorbent assay (ELISA). The serum concentration of IL1ß was initially increased within the first-hour post-challenge in Groups 2 and 3, followed by a significant decrease in concentration at 21d and 35d (p < 0.05) in Group 3. Only mild fluctuations in IL-6 occurred in group 2, as opposed to the 1.7-fold rapid increase in TNFα within 2 h post-challenge before decreasing at 6 h. An increase in pro-inflammatory cytokines was accompanied by an acute febrile response of 39.5 ± 0.38 °C (p < 0.05) at 2 h and 40.1 ± 0.29 °C (p < 0.05) at 4 h in Group 2 and Group 3, respectively. Serum testosterone decreased significantly (p < 0.05) in both treatment groups but remained significantly (p > 0.05) lower than in Group 1 throughout the study. There was a moderate negative association between testosterone and IL1ß (r = -0.473; p > 0.05) or TNFα (r = -0.527; p < 0.05) in Group 2. Serum LH also showed moderate negative associations with TNFα in Group 2 (r = -0.63; p < 0.05) and Group 3 (r = -0.54; p > 0.05). The results of this study demonstrated that M. haemolytica A2 and its OMP produced marked alterations in serum levels of pro-inflammatory cytokines and male reproductive hormones. The negative correlations between serum testosterone and inflammatory cytokines would suggest the potential role of OMP in causing male infertility by mediating innate inflammatory responses to suppress testosterone production in bucks.

Effect of Brachiaria decumbens as a novel supplementation on the production performance of broiler chickens.

The primary goal of this research was to elucidate the novel influence of Brachiaria decumbens supplementation on broiler chicken growth performance, nutritional digestibility, cecal microbiota, intestinal histomorphology, carcass characteristics, and meat quality. A total of 300 male day-old Ross 308 broiler chickens were randomly subjected to six different treatment groups having five replicates per treatment with 10 birds in each replicate. In treatment 1, broiler chickens were fed commercial diets with no added additives; in treatment 2, broiler chickens were offered commercial diets containing 100 mg/kg of the antibiotic oxytetracycline. However, in treatments 3, 4, 5, and 6, broiler chickens received similar commercial diets supplemented with 25 mg/kg, 50 mg/kg, 75 mg/kg, and 100 mg/kg of B. decumbens ground leaf powder, respectively, without antibiotics. Throughout the 42-day trial, the body weight gain and total feed intake for each replicate were recorded every week to determine the growth performance. Then, on 21th and 42nd day, ten broilers from each treatment (two in each replicate) were randomly selected and slaughtered to assess the digestibility of nutrients, histomorphology of the small intestine, the population of the cecal microbiota, carcass characteristics, as well as quality of both breast and drumstick muscle. There were differences (p < 0.05) in the growth performance, apparent ileal nutrient digestibility, intestinal histomorphology, carcass characteristics, and meat quality. Animals supplemented with 25 mg/kg of B. decumbens had higher growth performance and better ileal nutrient digestibility of protein, fiber, and ether extract, as well as higher villi height and carcass percentage with superior meat quality. Besides, the growth of E. faecalis during the starter phase was inhibited. In summary, B. decumbens supplementation at 25 mg/kg may be suggested as an optimal dose of as a prophylactic as well as an alternative for antibiotic growth promoter in stimulating the productivity of commercial broilers. This unconventional phytobiotic supplementation could be the key to replacing unnecessary antibiotics used in poultry feed.

Seminal and histopathological alterations in bucks challenged with Mannheimia haemolytica serotype a2 and its LPS endotoxin.

Pneumonic mannheimiosis is a widespread respiratory bacterial disease of small ruminants caused by Mannheimia haemolytica serotype A2. The disease is known to affect the respiratory organs of infected animals, but its effect on other vital and reproductive organs has not been fully explored. Previous studies have demonstrated increased serum pro-inflammatory cytokine concentration post-challenge with M. haemolytica A2 and its LPS, indicating systemic inflammation in the host. This study determined the potential tissue changes and alterations of sperm parameters due to infection of M. haemolytica A2 and its LPS endotoxin. In this study, twelve experimental bucks were randomly assigned to three groups of four bucks each: group 1 (control group) were intranasally inoculated with 2 mL of PBS pH 7.0, group 2 received 2 mL of 1.2 × 109 CFU/mL M. haemolytica A2 intranasally, and group 3 received 2 mL of LPS extracted from 1.2 × 109 CFU/mL of M. haemolytica A2 intravenously. Semen samples were collected at pre-determined intervals using an electro-ejaculator and analysed immediately after collection. All experimental bucks were slaughtered via exsanguination on day 60 to collect their vital and reproductive organs at necropsy, and the samples were processed and analysed for histopathological changes. The current study has revealed that bucks challenged with M. haemolytica A2 and its LPS exhibited alterations in semen parameters such as motility, wave pattern, viability, and morphological abnormalities. Mild to moderate histopathological changes of the lung, liver, testis, epididymis, vas deferens, prostate, and lymph nodes were also observed in both challenged groups. Therefore, this study revealed the potential harmful effects of respiratory mannheimiosis on the reproductive organs of the infected bucks and sheds light on the expanse of systemic effects of this disease.

Changes in selected cytokines, acute-phase proteins, gonadal hormones and reproductive organs of non-pregnant does challenged with Mannheimia haemolytica serotype A2 and its LPS endotoxin.

Previous investigations have revealed that lipopolysaccharide (LPS) endotoxin from certain Gram-negative bacteria could adversely affect the reproductive system of female animals. However, it is unknown whether LPS endotoxin of Mannheimia haemolytica serotype A2, the principal causative bacteria that cause pneumonic mannheimiosis in small ruminants, may also induce similar insidious effects. Therefore, this study aimed to investigate the effects of M. haemolytica serotype A2 and its LPS endotoxin on the responses of female gonadal hormones (progesterone and oestrogen), pro-inflammatory cytokines (interleukin-1ß, interleukin-6), acute-phase proteins (haptoglobin and serum amyloid A) and cellular changes via histopathology study of female reproductive organs of the treatment does. Twelve clinically healthy, non-pregnant, crossbred does were randomly allocated into three equal groups. Group 1 was administered intranasally with 2 ml of sterile phosphate-buffered saline (PBS) and served as a negative control group. Group 2 was challenged intranasally with 2 ml of bacterial inoculum containing 109 colony-forming units (CFU)/ml of M. haemolytica serotype A2, while Group 3 was challenged intravenously with 2 ml of LPS endotoxin extracted from 109 CFU/ml of M. haemolytica serotype A2. Following that, blood samples were collected serially at pre-determined intervals for serological analyses. All does were euthanised 60 days post-challenges, and tissue samples from the ovaries, oviducts, uterine horns, uterine body, cervix and vagina were collected for histopathological study. The serological result revealed a significant increase (p < 0.05) in the mean concentrations of progesterone, oestrogen, interleukin-1ß, interleukin-6, haptoglobin and serum amyloid A for both challenged groups. Histopathologically, all reproductive organs (except the cervix and vagina) from both challenged groups displayed significant cellular alterations (p < 0.05) characterised by haemorrhage and congestion, necrosis and degeneration, inflammatory cell infiltration and oedema. This study provides new information that elucidates the potential role of pneumonic mannheimiosis in the pathogenesis of female infertility amongst small ruminants.

Biomarkers, immune responses and cellular changes in vaccinated and non-vaccinated goats during experimental infection of M. haemolytica A2 under tropical conditions.

BACKGROUND: We investigated the biomarkers, immune responses and cellular changes in vaccinated and non-vaccinated goats experimentally challenged with M. haemolytica serotype A2 under rainy and hot tropical conditions. A total of twenty-four clinically healthy, non-pregnant, female goats randomly allocated to 2 groups of 12 goats each were used for the study. The 12 goats in each season were subdivided into three groups (n = 4), which served as the control (G-NEG), non-vaccinated (G-POS), and vaccinated (G-VACC). In week-1, the G-VACC received 2 mL of alum-precipitated pasteurellosis vaccine while G-POS and G-NEG received 2 ml of sterile PBS. In week 2, the G-POS and G-VACC received 1 mL intranasal spray containing 105 CFU of M. haemolytica serotype A2. Inoculation was followed by daily monitoring and weekly bleeding for eight weeks to collect data and serum for biomarkers and immune responses using commercial ELISA test kits. The goats were humanely euthanised at the end of the experiments to collect lungs and the submandibular lymph nodes tissue samples for gross and histopathological examinations. RESULTS: Regardless of the season, we have observed a significant (p < 0.05) increase in serum concentrations of acute-phase proteins (haptoglobin, serum amyloid A), proinflammatory cytokines (interleukine-1ß, interleukin-6), antibodies (immunoglobulin M, immunoglobulin G), and stress markers (cortisol and heat shock protein 70) in the G-POS goats compared to G-VACC and G-NEG. With regards to seasons, there was a significantly (p < 0.05) higher serum concentration with 1.5, 2 and 1-folds increase in the serum interleukin (IL)-1ß, cortisol, and heat shock protein (HSP)-70 in the G-POS during rainy compared to the hot season. Histopathology of the lungs in G-POS goats revealed inflammatory cell infiltration, degeneration, haemorrhage/congestion, and pulmonary oedema in the alveoli spaces; thickening of the interstitium, and desquamation of bronchiolar epithelium. Cellular changes in the lymph node were characterized by a marked hypercellularity in G-POS goats. CONCLUSION: Host responses to pneumonic mannheimiosis based on increased serum levels of biomarkers (cortisol, HSP70, IL-1ß and IL-6) and severe cellular changes seen in the lungs and lymph nodes of G-POS goats compared to vaccinated goats and control group are influenced by the high environmental humidity recorded in the rainy season. Increased relative humidity in the rainy season is a significant stress factor for the higher susceptibility and severity of pneumonic mannheimiosis of goats in the tropics. Vaccination of goats using the alum precipitated Pasteurella multocida vaccine before the onset of the rainy season is recommended to minimise mortality due to potential outbreaks of pneumonia during the rainy season.

The influence of feeding low and high level of Brachiaria decumbens diets on the hematology, serum biochemistry, and acute phase proteins of sheep.

The present study aims to determine the hematology, serum biochemistry, and acute phase proteins (APPs) responses of both serum and cerebrospinal fluid (CSF) in sheep fed with low and high levels of Brachiaria decumbens (B. decumbens) diets at different time phases. A total of 30 6-month-old male Dorper cross sheep were randomly divided into three treatment groups consisted of 10 sheep each. Treatment 1 (control) sheep were fed with Pennisetum purpureum and concentrates as the basal diet, whereas Treatments 2 and 3 sheep were fed with low (10%) and high (60%) level of B. decumbens, respectively. The hematology results revealed that there were significant differences (p < 0.05) in the red blood cells, mean corpuscular volume, mean corpuscular hemoglobin concentration, white blood cells, neutrophils, monocytes, eosinophils, basophils, platelets, and plasma proteins between groups. Except for packed cell volume, there were also significant differences in all hematology parameters at different time phases. All biochemistry parameters except creatinine revealed significant differences among treatment groups. However, there were significant differences in all parameters between time. On the other hand, APPs results showed significant differences in the serum haptoglobin and serum amyloid A in both serum and CSF between groups and time.

Responses of testosterone hormone and important inflammatory cytokines in bucks after challenge with Mannheimia haemolytica A2 and its LPS endotoxin.

Previous studies have shown that Mannheimia haemolytica A2 is the principal microorganism causing pneumonic mannheimiosis, a major bacterial respiratory disease among sheep and goats. The effect of this bacteria on the respiratory system is well-established. However, its effect on the reproductive physiology remains unclear. Therefore, this study aimed to determine the alterations in the level of pro-inflammatory cytokines and testosterone hormone post-inoculation with M. haemolytica serotype A2 and its lipopolysaccharide (LPS) endotoxin which were hypothesized to affect the reproductive functions of bucks. Twelve clinically healthy adult male goats were divided equally into three groups. Goats in group 1 were treated with 2 ml of sterile phosphate-buffered saline (PBS) pH 7.0 intranasally (negative control), group 2 with 2 ml of 109 colony-forming unit (CFU) of M. haemolytica serotype A2 intranasally (positive control), and group 3 were treated with 2 ml of lipopolysaccharide extracted from 109 CFU of M. haemolytica serotype A2 intravenously. Following inoculation, blood samples were collected via jugular venipuncture into plain tubes at pre-determined intervals for serum collection to determine the concentration of interleukin (IL)-1ß, IL6, tumor necrosis factor (TNF)-α, and testosterone hormone by using commercial ELISA test kits. Results from this study demonstrated that the inoculation of M. haemolytica A2 and its LPS increases the concentration of pro-inflammatory cytokines but decreases the concentration of testosterone hormone in challenged animals at most time points throughout the 56 days experimental period (p < 0.05). This study suggests that the M. haemolytica A2 and its LPS could alter the concentration of pro-inflammatory cytokines and testosterone hormone, which in turn, may negatively affect the reproductive functions of bucks.

Histopathological changes of reproductive organs of goats immunized with Corynebacterium pseudotuberculosis killed vaccine.

Caseous lymphadenitis (CLA) caused by Corynebacterium pseudotuberculosis is characterized by the development of abscesses, mainly in superficial and internal lymph nodes, visceral and reproductive organs in small ruminants. This study aims to examine the histopathological changes in reproductive organs of goats immunized with killed vaccine of C. pseudotuberculosis. In this study, twenty four (24) clinically healthy bucks and does were divided into four groups A, B, C and D. Animals in groups A and B were immunized with 0.5 and 1% formalin killed vaccine, respectively; followed by a booster dose. After the booster dose of immunization, groups A, B and C were challenged with C. pseudotuberculosis at 106 cfu/ml. Goats in group D were immunize and unchallenged and left as control group. All C. pseudotuberculosis infected animals were euthanized humanely 12 weeks post-challenged. Tissue samples such as testes, epididymis, spermatic cord, penis, pituitary gland, mammary gland, vulva, vagina, cervix, uterus, fallopian tube and ovaries were collected for histopathology study. Microscopic examination of all tissues (testes, seminiferous tubules, spermatic cord, penile tissues and the pituitary gland) in the male reproductive organs of the bucks that were inoculated with 2 ml of 0.5% and 1.0% of C. pseudotuberculosis killed vaccine showed normal (animals inoculated with 1.0%) to mild (animals inoculated with 0.5%) histopathological changes when compared with those from group C which showed varying degrees of histopathological changes (p < 0.01) in their various tissues. For the female does, similar histopathological changes were observed for the various tissues examined (ovaries, fallopian tubes, uterine horns, uterine tissues, cervix, vaginal, vulva, mammary glands and the pituitary glands) in which the vaccinated groups A &B showed a significantly (p < 0.001) less histopathological changes when compared with those in group C that showed varying degrees of histopathological changes in the reproductive organs investigated. This study showed the efficacy of C. pseudotuberculosis killed vaccine protecting against reproductive tissue damages cause by the active infection with the live bacteria in both bucks and does in the study area.

Responses of female reproductive hormones and histopathology in the reproductive organs and associated lymph nodes of Boer does challenged with Corynebacterium pseudotuberculosis and its immunogenic corynomycolic acid extract.

BACKGROUND: Corynebacterium pseudotuberculosis biotype ovis is a bacterium that causes caseous lymphadenitis (CLA), a chronic disease of sheep and goats characterized by the formation of suppurative abscesses in superficial and visceral lymph nodes and internal organs of small ruminants. This study was designed to evaluate the reproductive hormonal changes (estrogen and progesterone) and histopathology in the reproductive organs and associated lymph nodes of does challenged with C. pseudotuberculosis biotype ovis and its immunogen; corynomycolic acid. A total of 12 healthy non-pregnant female goats were grouped into three: A, B and C consisting of four does each. Group A was intradermally inoculated with 2 mL of sterile phosphate buffered saline (PBS) pH 7 (negative control group); group B was intradermally inoculated with 2 mL of corynomycolic acid extract (CMAs), while group C was intradermally inoculated with 2 mL of 109 colony-forming unit (cfu) of live C. pseudotuberculosis. Blood samples were also collected at predetermined intervals for estrogen and progesterone hormonal assays. The does were euthanized 90 days post challenge and tissue samples of the uterus, ovaries, fallopian tubes, cervix and associated lymph nodes were collected and fixed in 10% neutral buffered formalin for histopathological processing. The result showed various degrees of histopathological changes (hemorrhage, congestion, degeneration, necrosis, edema, leucocytic infiltrations) in the reproductive organs and associated lymph nodes of both inoculation groups. Increases in estrogen hormone concentration were observed in both inoculation groups in comparison to the control group. However, progesterone concentration was only increased in group C. This study highlighted that corynomycolic acid extract from C. pseudotuberculosis biotype ovis resulted in significant histopathology in the reproductive organs and associated lymph nodes of does and increase estrogen concentration.

Multiple gene targeting siRNAs for down regulation of Immediate Early-2 (Ie2) and DNA polymerase genes mediated inhibition of novel rat Cytomegalovirus (strain All-03).

BACKGROUND: Cytomegalovirus (CMV) is an opportunistic pathogen that causes severe complications in congenitally infected newborns and non-immunocompetent individuals. Developing an effective vaccine is a major public health priority and current drugs are fronting resistance and side effects on recipients. In the present study, with the aim of exploring new strategies to counteract CMV replication, several anti-CMV siRNAs targeting IE2 and DNA polymerase gene regions were characterized and used as in combinations for antiviral therapy. METHODS: The rat embryo fibroblast (REF) cells were transfected with multi siRNA before infecting with CMV strain ALL-03. Viral growth inhibition was measured by tissue culture infectious dose (TCID50), cytopathic effect (CPE) and droplet digital PCR (ddPCR) while IE2 and DNA polymerase gene knockdown was determined by real-time PCR. Ganciclovir was deployed as a control to benchmark the efficacy of antiviral activities of respective individual siRNAs. RESULTS: There was no significant cytotoxicity encountered for all the combinations of siRNAs on REF cells analyzed by MTT colorimetric assay (P > 0.05). Cytopathic effects (CPE) in cells infected by RCMV ALL-03 had developed significantly less and at much slower rate compared to control group. The expression of targeted genes was downregulated successfully resulted in significant reduction (P < 0.05) of viral mRNA and DNA copies (dpb + dpc: 79%, 68%; dpb + ie2b: 68%, 60%; dpb + dpc + ie2b: 48%, 42%). Flow cytometry analysis showed a greater percentage of viable and early apoptosis of combined siRNAs-treated cells compared to control group. Notably, the siRNAs targeting gene regions were sequenced and mutations were not encountered, thereby avoiding the formation of mutant with potential resistant viruses. CONCLUSIONS: In conclusion. The study demonstrated a tremendous promise of innovative approach with the deployment of combined siRNAs targeting at several genes simultaneously with the aim to control CMV replication in host cells.

Review of clinical aspects, epidemiology and diagnosis of haemotropic Mycoplasma ovis in small ruminants: current status and future perspectives in tropics focusing on Malaysia.

Mycoplasma ovis (formerly Eperythrozoon ovis) is an epierythrocytic parasitic bacterium of small ruminants known as haemotropic mycoplasma, which is transmitted mechanically by biting flies and contaminated instruments. Acute mycoplasmosis causes severe haemolytic anaemia and mortality in young animals. At the same time, chronic disease may produce mild anaemia and varying degrees of morbidity depending on several factors, including age, reproductive status, the plane of nutrition, immunological status and the presence of concurrent infection. Haemotropic Mycoplasma ovis is currently recognised as an emerging zoonotic pathogen which is widely distributed in the sheep and goat producing areas of tropics and subtropics, where the disease is nearly endemic. Human infection has been reported in pregnant women, immunocompromised patients and people exposed to animals and arthropods. The current diagnosis of haemoplasma relies on microscopic evaluation of Giemsa-stained blood smear and PCR. Although there are few published reports on the incidence of haemotropic Mycoplasma ovis infection of small ruminants in Malaysia, information on its prevalence, risk factors, severity and economic impacts is grossly inadequate. Therefore, a large-scale survey of small ruminant flocks is necessary to elucidate the current seroprevalence status and molecular characteristics of haemotropic M. ovis infection in Malaysia using ELISA and PCR sequencing technologies. In the future, surveillance programs, including vector forecast, quarantine, monitoring by periodic surveys and public enlightenment, will limit the internal and transboundary spread of M. ovis, enhance control efforts and mitigate production losses in Malaysia.

Establishment of lung auscultation scoring method and responses of acute phase proteins and heat shock proteins in vaccinated and non-vaccinated goats.

Pneumonic pasteurellosis is an economically important infectious disease in the small ruminant industry which causes sudden death and loss for farmers. Nonetheless, this disease is still a common sight in sheep and goats in Malaysia, probably due to the unpopular usage of pasteurellosis vaccine or inappropriate vaccination practices. The aim of this study was designed to classify the severity of pneumonia via the establishment of auscultation scoring method and to quantify the acute phase proteins and heat shock proteins responses from vaccinated and non-vaccinated goats. Goat farms, consist of vaccinated and non-vaccinated farms, were selected in this study: where 15 clinically normal healthy goats and 9 pneumonic goats were selected from vaccinated farms whereas 15 clinically normal healthy goats and 31 pneumonic goats from non-vaccinated farms were selected for this study. Crackle lung sounds were not detected in both vaccinated and non-vaccinated normal goats. However, vaccinated pneumonic goats showed mild crackle lung sound while non-vaccinated pneumonic goats exhibited moderate crackle lung sound. There were significant increases (p < 0.05) in acute phase proteins and heat shock proteins concentrations for the non-vaccinated pneumonic goats group. In this study, conclusion can be made that the vaccinated goats exhibited very mild clinical responses of pneumonia and non-significant biomarker responses compared to the non-vaccinated goats. Thus, vaccination is an effective preventive measure to control pneumonic pasteurellosis and acute phase proteins and heat shock proteins can be considered as future biomarkers in screening and rapid diagnostic method for this particular disease.

Responses of pro-inflammatory cytokines, acute phase proteins and cytological analysis in serum and cerebrospinal fluid during haemorrhagic septicaemia infection in buffaloes.

Sudden death is usually the main finding in field animals during haemorrhagic septicaemia outbreaks caused by Pasteurella multocida type B:2 that causes acute, fatal and septicaemic disease in cattle and buffaloes. This situation may be due to failure in early detection of the disease where early treatment of antibiotics may improve the prognosis of the animal and other surviving animals. Thus, there is a grey area on the knowledge on the potential usage of pro-inflammatory cytokines and acute phase proteins as early biomarkers in the diagnosis of haemorrhagic septicaemia. In addition, exploration of the cerebrospinal fluid during infection has never been studied before. Therefore, this study was designed to fill up the grey areas in haemorrhagic septicaemia research. Twenty-one buffalo calves were divided into seven treatment groups where group 1 was inoculated orally with 10 mL of sterile phosphate-buffered saline pH 7 which act as a negative control group. Groups 2 and 3 were inoculated orally and subcutaneously with 10 mL of 1012 colony-forming unit of P. multocida type B:2. Group 4 and 5 buffaloes were inoculated orally and intravenously with 10 mL of lipopolysaccharide broth. Groups 6 and 7 were administered orally and subcutaneously with 10 mL of outer membrane protein broth. During the post-infection period of 21 days, blood and cerebrospinal fluid were sampled for the analyses of pro-inflammatory cytokines, acute phase proteins and cytological examination. Buffalo calves infected with P. multocida and its immunogens via different routes of inoculation showed significant changes (p < 0.05) of pro-inflammatory cytokines, acute phase proteins and cytological changes in both the serum and cerebrospinal fluid. Buffalo calves from groups 3 and 7 showed the highest pro-inflammatory cytokines, whereas group 6 had the highest acute phase protein concentration and group 5 revealed the highest value for cytology changes. In summary, results obtained in this study could be used as a profiling study to add novel knowledge to the haemorrhagic septicaemia research as well as the development of biomarkers.

The re-emerging of orf virus infection: A call for surveillance, vaccination and effective control measures.

Orf disease is known to be enzootic among small ruminants in Asia, Africa, and some other parts of the world. The disease caused by orf virus is highly contagious among small ruminant species. Unfortunately, it has been neglected for decades because of the general belief that it only causes a self-limiting disease. On the other hand, in the past it has been reported to cause huge cumulative financial losses in livestock farming. Orf disease is characterized by localized proliferative and persistent skin nodule lesions that can be classified into three forms: generalized, labial and mammary or genitals. It can manifest as benign or malignant types. The later type of orf can remain persistent, often fatal and usually causes a serious outbreak among small ruminant population. Morbidity and mortality rates of orf are higher especially in newly infected kids and lambs. Application of antibiotics together with antipyretic and/or analgesic is highly recommended as a supportive disease management strategy for prevention of subsequent secondary microbial invasion. The presence of various exotic orf virus strains of different origin has been reported in many countries mostly due to poorly controlled cross-border virus transmission. There have been several efforts to develop orf virus vaccines and it was with variable success. The use of conventional vaccines to control orf is a debatable topic due to the concern of short term immunity development. Following re-infection in previously vaccinated animals, it is uncommon to observe the farms involved to experience rapid virus spread and disease outbreak. Meanwhile, cases of zoonosis from infected animals to animal handler are not uncommon. Despite failures to contain the spread of orf virus by the use of conventional vaccines, vaccination of animals with live orf virus is still considered as one of the best choice. The review herein described pertinent issues with regard to the development and use of potential effective vaccines as a control measure against orf virus infection.

Can treatment of Brachiaria decumbens (signal grass) improve its utilisation in the diet in small ruminants?-a review.

Brachiaria decumbens is an extremely productive tropical grass due to its aggressive growth habit and its adaptation to a varied range of soil types and environments. As a result of the vast availability, treated B. decumbens demonstrates as a promising local material that could be utilised as an improved diet for sheep and goats. Despite the fact that the grass significantly increases weight gains in grazing farm animals, there were many reports of general ill-thrift and sporadic outbreaks of photosensitivity in livestock due to the toxic compound of steroidal saponin found in B. decumbens. Ensiling and haymaking were found to be effective in removing toxin and undesirable compounds in the grass. Biological treatments using urea, activated charcoal, polyethylene glycol, and effective microorganisms were found to be useful in anti-nutritional factor deactivation and improving the nutritive values of feedstuffs. Besides, oral administration of phenobarbitone showed some degree of protection in sheep that fed on B. decumbens pasture. In this review, we aim to determine the effect of B. decumbens toxicity and possible treatment methods on the grass to be used as an improved diet for small ruminant.

Reproductive hormonal variations and adenohypophyseal lesions in pre-pubertal buffalo heifers inoculated with Pasteurella multocida type B: 2 and its immunogens.

BACKGROUND: Hemorrhagic septicemia is a fatal disease of cattle and buffaloes caused by P. multocida. Although the pathogenesis of the bacteria has been well established in literature, there is a paucity of information on the possible role of the bacteria and its immunogens; lipopolysaccharide (LPS) and outer membrane proteins (OMPs) on the reproductive capacity of buffalo heifers. METHODS: In this study, twenty one healthy prepubertal female buffaloes aged 8 months were divided into seven groups of 3 buffaloes each (G1-G7). Group 1 (G1) served as the negative control group and were inoculated orally with 10 mL sterile Phosphate Buffer Saline (PBS), groups 2 (G2) and 3 (G3) were inoculated orally and subcutaneously with 10 mL of 1012 colony forming unit (cfu) of P.multocida type B: 2, while groups 4 (G4) and 5 (G5) received 10 mL of bacterial LPS orally and intravenously, respectively. Lastly, groups 6 (G6) and 7 (G7) were orally and subcutaneously inoculated with 10 mL of bacterial OMPs. Whole blood was collected in EDTA vials at stipulated time points (0, 2, 4, 6, 8, 10, 12, 24, 36, 48, 72, 120, 168, 216, 264, 312, 360, 408, 456 and 504 h), while tissue sections of the pituitary glands were collected and transported to the histopathology laboratory in 10% buffered formalin for processing and Hematoxylin and eosin staining. Plasma levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), progesterone (PG), estradiol (EST) and gonadotrophin releasing hormone (GnRH) were determined. RESULTS: The histopathological lesions observed in the pituitary gland included hemorrhage, congestion, inflammatory cell infiltration, hydropic degeneration, necrosis and edema. These changes were higher (p < 0.05) in distribution and severity in G3, G6 and G7. Hormonal concentrations of LH, FSH, PG, EST and GnRH declined in all inoculation groups as time elapsed and were lower (p < 0.05) than that of the control group. CONCLUSION: Based on these findings, P.multocida B: 2 and its immunogens can be said to negatively affect the hypothalamic-pituitary-gonadal axis, resulting in decreased levels of reproductive hormones which may predispose to infertility in buffalo heifers.

Involvement of the nervous system following experimental infection with Pasteurella multocida B:2 in buffalo (Bubalus bubalis): A clinicopathological study.

Haemorrhagic septicaemia (HS) is an acute, fatal, septicaemic disease of cattle and buffaloes caused by one of two specific serotypes of Pasteurella multocida B:2 and E:2 in Asian and African, respectively. It is well known that HS affect mainly the respiratory and digestive tracts. However, involvement of the nervous system in pathogenesis of HS has been reported in previous studies without details. In this study, nine buffalo calves of 8 months old were distributed into three groups. Animals of Group 1 and 2 were inoculated orally and subcutaneously with 10 ml of 1 × 10(12) cfu/ml of P. multocida B:2, respectively, while animals of Group 3 were inoculated orally with 10 ml of phosphate buffer saline as a control. All calves in Group 1 and Group 3 were euthanised after 504 h (21 day) post-infection, while calves in Group 2 had to euthanise after 12 h post-infection as they develop sever clinical signs of HS. Significant differences were found in Group 2 in the mean scores of clinical signs, gross and histopathological changes which mainly affect different anatomic regions of the nervous system. In addition, successful bacterial isolation of P. multocida B:2 were obtained from different sites of the nervous system. On the other hand, less sever, clinical, gross and histopathological changes were found in Group 1. These results provide for the first time strong evidence of involving of the nervous system in pathogenesis of HS, especially in the peracute stage of the disease.

New sites of localisation of Pasteurella multocida B:2 in buffalo surviving experimental haemorrhagic septicaemia.

BACKGROUND: Haemorrhagic septicaemia (HS) is an acute septicaemic disease of buffalo and cattle caused by Pasteurella multocida B:2 and E:2. Field outbreaks of HS are known to result in localisation of bacteria in the tonsils of surviving buffalo, confirming that animals can become carriers and the role of respiratory tract in the transmission of the disease. This report describes additional sites of localisation of P. multocida B:2 in surviving buffalo following experimental induction of HS. RESULTS: Following P. multocida B:2 infection, all calves in group 1 and one calf in group 2 that was allowed to commingle with infected calves from group 1 were euthanised within 48 h. Pasteurella multocida B:2 was detected from the nasal and rectal swab samples on days 5 and 6 from the remaining calves in group 2. The first injection of dexamethasone into the carrier animals resulted in reemergence in samples from the nose, rectum and vagina. However, subsequent dexamethasone injections failed to re-activate P. multocida B:2. When surviving carrier calves in group 2 were euthanised at the end of the experiment, P. multocida B:2 was detected in the lungs and various organs of the respiratory, gastrointestinal and urinary tracts. CONCLUSIONS: Commingling naive buffalo calves with calves acutely infected with P. multocida B:2 resulted in carriers among surviving buffalo. Pasteurella was found in various organs of the respiratory, gastrointestinal and urinary tracts, suggesting their role in the pathogenesis of HS.

Digestibility, Growth Performance, Body Measurement and Hormone of Sheep Fed with Different Levels of Brachiaria decumbens Diets.

Limited data are available regarding the effects of Brachiaria decumbens on sheep's growth performance at different times. Therefore, this current study focused on sheep's nutrient apparent digestibility, feed efficiency, body index, and growth hormone when they are fed with low and high levels of B. decumbens diets. A total of 30 six-month-old male Dorper cross sheep were divided randomly into three treatment groups with 10 sheep per treatment. Treatment 1 (control) sheep were fed with Pennisetum purpureum and pellets as the basal diet, whereas Treatment 2 and 3 sheep were fed with feed mixed with low (10%) and high (60%) levels of B. decumbens, respectively. The study was conducted in two phases consisting of short-term feeding (seven days) and long-term feeding (90 days). Throughout the experiment, daily fecal voided were collected in the morning for seven days continuous before the end of each feeding phases for the determination of nutrient apparent digestibility. The amount of feed offered and refusals plus body weight gain were recorded daily to determine the feed efficiency (FE). Besides, the body measurements of each sheep from every treatment were measured weekly and blood samples were collected for the analysis of growth hormone (GH) concentration. There were significant differences (p < 0.05) in the nutrient apparent digestibility, growth performance, body measurement, and GH concentration among treatment sheep throughout the study period. Treatment 3 sheep fed with 60% of B. decumbens diet revealed the lowest dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), and acid detergent fiber (ADF) digestibility during the long-term feeding. Likewise, Treatment 3 (T3) sheep had the lowest total bodyweight gain, average daily gain, total feed intake, and daily feed intake among treatment sheep. The heart girth index (HGI) of T3 sheep was also significantly lower during the short-term feeding. Moreover, the GH concentration of T3 sheep was significantly lower as compared to the control that decreases steadily throughout the study period. In conclusion, high levels of B. decumbens showed the most significant results out of all three treatments indicating the presence of saponins, which produce negative effects on the sheep's overall performance.

Data tersedia mengenai kesan jumlah Brachiaria decumbens yang berbeza terhadap prestasi pertumbuhan biri-biri pada tempoh masa yang berbeza adalah terhad. Oleh itu, kajian semasa ini memberi tumpuan kepada kebolehcernaan ketara nutrien, kecekapan makanan, indeks badan, dan hormon pertumbuhan bebiri yang diberi makan dengan diet B. decumbens pada tahap rendah dan tinggi. Sebanyak 30 ekor bebiri Dorper jantan berumur 6 bulan dibahagikan secara rawak kepada 3 kumpulan rawatan dengan 10 ekor bebiri setiap kumpulan. Rawatan 1 (kawalan) bebiri diberi makan dengan Pennisetum purpureum dan pelet sebagai diet asas, manakala bebiri Rawatan 2 dan 3 masing-masing diberi makan dengan campuran B. decumbens pada tahap rendah (10%) dan tinggi (60%). Kajian ini dijalankan dalam dua fasa yang terdiri daripada pemakanan jangka pendek (7 hari) dan panjang (90 hari). Sepanjang eksperimen, najis harian yang dibuang dikumpulkan pada waktu pagi selama 7 hari berterusan sebelum tamat setiap fasa pemakanan untuk menentukan kebolehcernaan ketara nutrien. Jumlah makanan yang diberi, sisa makanan, dan penambahan berat badan direkodkan setiap hari untuk menentukan kecekapan makanan (FE). Selain itu, ukuran badan setiap bebiri daripada setiap rawatan diukur setiap minggu serta sampel darah diambil untuk analisis kepekatan hormon pertumbuhan (GH). Terdapat perbezaan yang signifikan (p < 0.05) dalam kebolehcernaan ketara nutrien, prestasi pertumbuhan, ukuran badan, dan kepekatan GH di kalangan bebiri rawatan sepanjang tempoh kajian. Bebiri rawatan 3 (T3) yang diberi makan dengan 60% diet B. decumbens menunjukkan kebolehcernaan bahan kering (DM), protein kasar (CP), gentian detergen neutral (NDF), dan gentian detergen asid (ADF) yang paling rendah semasa pemberian makanan jangka panjang. Bebiri T3 mempunyai jumlah penambahan berat badan, purata kenaikan harian, jumlah pengambilan makanan, dan pengambilan makanan harian yang paling rendah di kalangan bebiri rawatan. Indeks lilitan jantung (HGI) bebiri T3 juga jauh lebih rendah semasa pemberian makanan jangka pendek. Selain itu, kepekatan GH bebiri T3 adalah jauh lebih rendah berbanding dengan bebiri kawalan yang menurun secara berterusan sepanjang tempoh kajian. Kesimpulannya, tahap B. decumbens yang tinggi menunjukkan keputusan yang paling ketara daripada ketiga rawatan yang menunjukkan kehadiran saponins boleh menghasilkan kesan negatif terhadap prestasi keseluruhan bebiri.