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.

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.

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.

Genome-Wide Analysis and Molecular Characterization of Orf Virus Strain UPM/HSN-20 Isolated From Goat in Malaysia.

Contagious ecthyma commonly known as Orf is a globally important, highly contagious zoonotic, transboundary disease that affects domestic and wild ruminants. The disease is of great economic significance causing an immense impact on animal health, welfare, productivity, and trade. Detailed analysis of the viral genome is crucial to further elucidate the molecular mechanism of Orf virus (ORFV) pathogenesis. In the present study, a confluent monolayer of lamb testicle cells was infected with the processed scab sample obtained from an infected goat. The presence of the virus was confirmed using polymerase chain reaction and electron microscopy, while its genome was sequenced using next-generation sequencing technology. The genome sequence of Malaysian ORFV strain UPM/HSN-20 was found to contain 132,124 bp with a G + C content of 63.7%. The homology analysis indicates that UPM/HSN-20 has a high level of identity 97.3-99.0% with the other reference ORFV strain. Phylogenetic analysis revealed that ORFV strain UPM/HSN-20 is genetically more closely related to ORFV strain XY and NP from China. The availability of the genome-wide analysis of ORFV UPM/HSN-20 strain from Malaysia will serve as a good platform for further understanding of genetic diversity, ORFV infection, and strategic development for control measures.

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.

Edible bird's nest protects histomorphology of rat's uterus against cadmium (Cd) toxicity through a reduction of Cd deposition and enhanced antioxidant activity.

Cadmium (Cd) is often associated with reproductive disorders of mammals. Edible bird's nest (EBN) is a natural food product made of swiftlet's salivary secretion used to make their nests and it has been consumed as a tonic food for decades. This research aimed to study the protective effects of EBN against Cd-induced uterine toxicity in Sprague Dawley rats. Thirty (30) female Sprague Dawley rats were assigned into five groups as follows: group 1- negative control (NC) received distilled water; group 2 - positive control (PC) administered with CdCl2, 5 mg/kg BW; while groups EBN-1, EBN-2, and EBN-3 received CdCl2 (5 mg/kg BW) plus graded concentrations of 60, 90 and 120 mg/kg BW of EBN, respectively. After four weeks of daily oral treatment, rats were euthanized to collect the uterus for evluations of histopathological changes, Cd concentrations and Metallothionein (MT) expressions using H&E stain, inductive coupled plasma mass spectrometry (ICP-MS) and immunohistochemistry, respectively. Blood samples were collected for superoxide dismutase (SOD) analysis using SOD assay kit. Results revealed that the CdCl2 without EBN supplement (PC) group had elevated levels of Cd in the uterus along with increased MT expressions and decreased SOD enzyme activity as compared to the NC group. Moreover, uterine histopathological changes, including glandular cysts and loss of normal structure of luminal epithelium (LE) and glandular epithelium (GE) were found in the PC group. Interestingly, groups treated with CdCl2 along with EBN (EBN1, EBN2, EBN3) showed lower levels of uterine tissue Cd deposition and MT expression, lower degenerative changes with normal histomorphology of glands, and increased SOD activity as compared to the PC group. Overall, the findings revealed that oral exposure to Cd at a dose of 5 mg/kg BW resulted in significant alterations in the rat's uterus. However, the toxicity effect was averted by EBN treatment in a dose dependant manner; highest protection achieved with EBN 120 mg/kg BW, through a possible detoxification mechanism and prevention of Cd deposition.

Mycobacterium Tuberculosis and Avium Complex Investigation among Malaysian Free-Ranging Wild Boar and Wild Macaques at Wildlife-Livestock-Human Interface.

Wild animals are considered reservoirs, contributing to the transmission of emerging zoonotic diseases such as tuberculosis (TB). A cross-sectional study was conducted by opportunistic sampling from fresh carcasses of free-ranging wild boar (n = 30), and free-ranging wild macaques (n = 42). Stained smears from these tissues were tested for acid-fast bacilli (AFB) with Ziehl-Neelsen staining. Mycobacterial culture was conducted using Lowenstein-Jensen media and Middlebrook 7H11 agar media. Polymerase chain reaction (PCR) was performed through the detection of the 16S rRNA gene, with multiple sets of primers for the detection of Mycobacterium tuberculosis complex (MTBC) and Mycobacterium avium complex (MAC). In wild boars, 30% (9/30; 95% Confidence Interval: 16.7-47.9%) of examined samples showed gross tuberculosis-like lesions (TBLLs). Multiple nodular lesions that were necrotic/miliary with cavitation were found in the submandibular lymph nodes, tonsils, lungs, kidney and liver, while single nodular lesions were found in the mediastinal lymph nodes, spleen and mesenteric lymph nodes. Conventional PCR on the submandibular lymphoid tissues of wild boar (nine samples with TBLLs and three non-TBLL samples) showed that 75% (9/12) were positive for Mycobacterium bovis (95% CI: 46.8-91.1), and 91% (CI: 64.6-98.5) were positive for Mycobacterium avium. For macaques, 33.3% (10/30) were positive for M. avium (95% CI: 19.2-51.2) but negative for MTBC.

Immunomodulatory Strategies for Parapoxvirus: Current Status and Future Approaches for the Development of Vaccines against Orf Virus Infection.

Orf virus (ORFV), the prototype species of the parapoxvirus genus, is the causative agent of contagious ecthyma, an extremely devastating skin disease of sheep, goats, and humans that causes enormous economic losses in livestock production. ORFV is known for its ability to repeatedly infect both previously infected and vaccinated sheep due to several immunomodulatory genes encoded by the virus that temporarily suppress host immunity. Therefore, the development of novel, safe and effective vaccines against ORFV infection is an important priority. Although, the commercially licensed live-attenuated vaccines have provided partial protection against ORFV infections, the attenuated viruses have been associated with major safety concerns. In addition to safety issues, the persistent reinfection of vaccinated animals warrants the need to investigate several factors that may affect vaccine efficacy. Perhaps, the reason for the failure of the vaccine is due to the long-term adaptation of the virus in tissue culture. In recent years, the development of vaccines against ORFV infection has achieved great success due to technological advances in recombinant DNA technologies, which have opened a pathway for the development of vaccine candidates that elicit robust immunity. In this review, we present current knowledge on immune responses elicited by ORFV, with particular attention to the effects of the viral immunomodulators on the host immune system. We also discuss the implications of strain variation for the development of rational vaccines. Finally, the review will also aim to demonstrate future strategies for the development of safe and efficient vaccines against ORFV infections.

Detection of Mycobacterium tuberculosis complex antibodies in free-ranged wild boar and wild macaques in selected districts in Selangor and reevaluation of tuberculosis serodetection in captive Asian elephants in Pahang, Peninsular Malaysia.

Tuberculosis (TB) is a chronic inflammatory and zoonotic disease caused by Mycobacterium tuberculosis complex (MTBC) members, affecting several domestic animals, wildlife species and humans. The preliminary investigation was aimed to detect antibody against MTBC among indigenous wildlife which are free-ranged wild boar, free-ranged wild macaques and captive Asian elephants in selected areas of Selangor and elephant conservation centre in Pahang, respectively. The results indicate that MTBC serodetection rate in wild boar was 16.7% (7.3-33.5 at 95% confidence interval (CI)) using an in-house ELISA bPPD IgG and 10% (3.5-25.6 at 95% CI) by DPP®VetTB assay, while the wild macaques and Asian elephant were seronegative. The univariate analysis indicates no statistically significant difference in risk factors for sex and age of wild boar but there was a significant positive correlation (P<0.05) between bovine TB in dairy cattle and wild boar seropositivity in the Sepang district.

Further Insights Into Caprine Arthritis Encephalitis (CAE): The Current Status of Seroprevalence Among Small Ruminants in Two Selected States of Peninsular Malaysia.

Caprine arthritis-encephalitis virus (CAEV) is a member of the genus lentivirus causing caprine arthritis-encephalitis (CAE), a chronic inflammatory condition affecting the lungs, joints, udder and central nervous system of small ruminants such as sheep and goats. CAE is distributed worldwide and is recognised as a significant cause of morbidity and decreased milk production in dairy goats. Earlier studies highlighted the clinicopathological features and supplied preliminary serological evidence for the existence of CAE among selected goat herds in Malaysia. Therefore, this study aims to provide further insights into the seroprevalence and contributing factors of CAE among sheep and goat herds in two states of Peninsular Malaysia. The blood samples and biodata were randomly collected from a total of 262 individual sheep (40) and goat (222) in seven smallholder farms. Blood sera were tested for specific anti-CAEV antibodies using Qayee-Bio CAEV sandwich-ELISA test kits according to standard procedures. Our results of the study revealed 21.4% (95% CI: 15.8-28.6) apparent and 20.6% (95% CI: 14.5-27.8) true seroprevalence with significant differences (p < 0.05) in seroconversion rates between the states, farms, production systems and breeds of small ruminants. The prevalence of CAE in the Malaysian Peninsular is a potential threat to the small ruminant industry and developing agricultural economy. Further studies are required to determine the genetic characteristics, distribution and risk factors of CAEV for effective prevention and control in Malaysia.

Virus Kaprin Artritis Ensefalitis (CAEV) merupakan ahli kumpulan dalam genus virus lentivirus dimana akan menyebabkan penyakit kaprin artritis ensefalitis (CAE) di mana penyakit ini akan menyebabkan keradangan kronik pada paru-paru, sendi, kelenjar mamari dan sistem saraf pusat bagi haiwan ruminan kecil seperti bebiri dan kambing. CAE telah merebak ke seluruh dunia dan penyakit ini akan menyebabkan penularan wabak pada kadar morbiditi yang tinggi dan mengurangkan kuantiti penghasilan susu bagi kambing tenusu. Kajian terdahulu memberi penekanan kepada dapatan klinikal patologi dan data bukti serologi kewujudan penyakit CAE dalam kalangan gerompok kambing di Malaysia. Maka, kajian kini bertujuan memberikan pendedahan awal berkaitan kadar kelaziman serologi dan faktor yang menyumbang penyakit CAE dalam kalangan bebiri dan kambing bagi dua negeri di Semanjung Malaysia. Sampel darah dan data biologi telah dikumpulkan secara rawak dengan jumlah sampel 262 ekor (40 ekor bebiri dan 222 ekor kambing) daripada tujuh buah ladang peternak kecil. Serum yang telah dikumpul diuji dengan antibodi spesifik anti-CAEV dengan menggunakan prosedur piawai kit elisa daripada Qayee-Bio CAEV. Keputusan kajian ini menunjukkan kadar kelaziman serologi 21.4% (95% CI: 15.8­28.6) jelas dan 20.6% (95% CI: 14.5­27.8) kadar kelaziman benar dengan perbezaan ketara (p < 0.05) dalam kadar perubahan kelaziman serologi antara negeri, ladang, sistem produksi dan baka haiwan ruminan kecil. Dengan wujudnya kelaziman penyakit CAE di Semenanjung Malaysia ini akan menyumbang kepada kemungkinan ancaman negatif terhadap industri ruminan kecil dan sektor ekonomi dalam bidang penternakan. Lebih banyak kajian diperlukan untuk menentukan ciri genetik virus penyebab penyakit ini, taburan penyakit dan faktor penyumbang bagi CAEV supaya dapat mengadakan kawalan dan pencegahan efektif bagi penyakit ini di Malaysia.

Can Yucca schidigera Be Used to Enhance the Growth Performance, Nutrient Digestibility, Gut Histomorphology, Cecal Microflora, Carcass Characteristic, and Meat Quality of Commercial Broilers Raised under Tropical Conditions?

This study aims to study the effect of Yucca shidigera as a phytobiotic supplementation in enhancing the production performance of commercial broilers reared under tropical environments. A total of 300 male day-old Ross 308 broiler chicks were randomly allocated into six treatment groups. Treatment 1 broilers were fed with commercial diets without antibiotics. Treatment 2 broilers were fed with commercial diets added with 100 mg/kg oxytetracycline antibiotic. Treatment 3, 4, 5, and 6 were fed with the same commercial diets added with 25, 50, 75, and 100 mg/kg Y. shidigera, respectively, without antibiotic. Throughout the six weeks study period, body weight and feed intake were recorded weekly for each replicate to calculate the body weight gain and feed conversion ratio. In addition, the nutrient digestibility, gut histomorphology, cecal microflora population, carcass characteristics, and meat quality were determined. The results showed significant differences (p < 0.05) in the growth performance, apparent ileal nutrient digestibility, gut histomorphology, carcass traits, and meat quality. Overall, T6 broilers supplemented with 100 mg/kg Y. shidigera demonstrated the best production performances as compared to the other treatment broilers. In summary, information from this study will be valuable for the usability of Y. schidigera, which could be developed as a feed additive to replace antibiotics in the poultry sector in the tropics.

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.

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.

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.

Immediately early 2 (IE-2) and DNA polymerase SiRNA as virus-specific antiviral against novel transplacental cytomegalovirus strain ALL-03 in vitro.

OBJECTIVE: This study investigated the suitability of siRNA targeting specific genes that cause inhibition of virus replication in vitro especially for the virus that capable of crossing placenta and we employed a novel transplacental rat cytomegalovirus that mimics infection in human. METHODS: Six unique siRNAs with three each targeting different regions of IE2 (ie2a, ie2b and ie2c) and DNA polymerase (dpa, dpb and dpc) were prepared and tested for antiviral activities. The efficacy as an antiviral was determined in in-vitro by measuring TCID50 virus titer, severity of virus-induced cytopathic effect (CPE), intracellular viral genome loads by droplet digital PCR, the degree of apoptosis in siRNA-treated cells and relative expression of viral mRNA in infected Rat Embryo Fibroblast (REF) cells. FINDINGS: Remarkably, the siRNAs: dpa, dpb and IE2b, significantly reduced virus yield (approximately >90%) compared to control group at day 18 post infection (p.i). Changes in CPE indicated that DNA polymerase siRNAs were capable of protecting cells against CMV infection at day 14 p.i with higher efficiency than GCV (at the concentration of 300 pmol). Gene expression analysis revealed a marked down regulation of the targeted DNA polymerase gene (73.9%, 96.0% and 90.7% for dpa, dpb and dpc siRNA, respectively) and IE2 gene (50.8%, 49.9% and 15.8% for ie2a, ie2b and ie2c siRNA, respectively) when measured by RT-qPCR. Intracellular viral DNA loads showed a significant reduction for all the DNA polymerase siRNAs (dpa: 96%, dpb: 98% and dpc:92) compared to control group (P < 0.05). CONCLUSION: In conclusion, this study clearly highlighted the feasibility of RNAi as an alternative antiviral therapy that could lead to controlling the CMV infection.

Risk Factors and Severity of Gastrointestinal Parasites in Selected Small Ruminants from Malaysia.

The productivity of smallholder sheep and goat flocks is constrained by high morbidity and mortality of young stock due to helminthosis and coccidiosis. We hypothesized that gastrointestinal parasites are prevalent and may cause severe infections amongst small ruminants in Malaysia. A cross-sectional survey was conducted between March and December 2019 to investigate the prevalence, risk factors, and levels of infection with gastrointestinal strongyle and coccidia in selected smallholder goat flocks in Negeri Sembilan, Malaysia. A total of 257 blood and fecal samples and management data were collected from four farms in Negeri Sembilan. Gastrointestinal parasites were detected by routine sodium chloride floatation, and the McMaster technique was used to quantify the fecal eggs/oocysts per gram outputs (EPG/OPG). The severity of infection was classified as mild (50-799), moderate (800-1200), or severe (>1200). The packed cell volume (PCV) was determined by microhematocrit centrifugation and classified as anemic or non-anemic. Coprological examination revealed an overall prevalence of 78.6% (CI = 72.74-83.44) and 100% flock level prevalence of strongyle and coccidia infection among goats from Negeri Sembilan with a higher infection in flock A-Lenggeng (95.6%) than B-Senawang (87.3%), D-Mendom (80.6%), or C-Seremban (60.0%). The co-infections of strongyle + Eimeria (50.6; CI = 44.50 to 56.64) were more common than single infections of either strongyle (16.7%; CI = 12.66 to 21.78) or Eimeria (4.3%; CI = 2.41 to 7.50). Quantitative analysis has revealed different (p < 0.05) patterns of EPG/OPG in various categories of goats. In total, there were 49.8% mild, 8.6% moderate, and 13.6% severe infections of strongyle and 40.1% mild, 6.6% moderate, and 19.8% severe infections of coccidia among goats. The mean PCV of goats with severe strongyle infection (24.60 ± 0.85) was significantly (p < 0.05) lower than the moderate (26.90 ± 1.15), or mild (28.23 ± 0.50) infections and the uninfected (30.4 ± 0.71). There were increased odds of infection with strongyle and coccidia among female (OR = 3.2) and adult (OR = 11.0) goats from smallholder flocks in Negeri Sembilan. In conclusion, gastrointestinal strongyles and coccidia occur at high frequency among smallholder goats, and there is a higher risk of infection amongst the adult and female stock.

Mycobacterium tuberculosis complex in wildlife: Review of current applications of antemortem and postmortem diagnosis.

Tuberculosis (TB) is a chronic inflammatory and zoonotic disease caused by Mycobacterium tuberculosis complex (MTBC) members, which affects various domestic animals, wildlife, and humans. Some wild animals serve as reservoir hosts in the transmission and epidemiology of the disease. Therefore, the monitoring and surveillance of both wild and domestic hosts are critical for prevention and control strategies. For TB diagnosis, the single intradermal tuberculin test or the single comparative intradermal tuberculin test, and the gamma-interferon test, which is regarded as an ancillary test, are used. Postmortem examination can identify granulomatous lesions compatible with a diagnosis of TB. In contrast, smears of the lesions can be stained for acid-fast bacilli, and samples of the affected organs can be subjected to histopathological analyses. Culture is the gold standard test for isolating mycobacterial bacilli because it has high sensitivity and specificity compared with other methods. Serology for antibody detection allows the testing of many samples simply, rapidly, and inexpensively, and the protocol can be standardized in different laboratories. Molecular biological analyses are also applicable to trace the epidemiology of the disease. In conclusion, reviewing the various techniques used in MTBC diagnosis can help establish guidelines for researchers when choosing a particular diagnostic method depending on the situation at hand, be it disease outbreaks in wildlife or for epidemiological studies. This is because a good understanding of various diagnostic techniques will aid in monitoring and managing emerging pandemic threats of infectious diseases from wildlife and also preventing the potential spread of zoonotic TB to livestock and humans. This review aimed to provide up-to-date information on different techniques used for diagnosing TB at the interfaces between wildlife, livestock, and humans.

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.

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.