Abdul Aziz Saharee

Reproductive Pathological Changes Associated with Experimental Subchronic Corynebacterium pseudotuberculosis Infection in Nonpregnant Boer Does

Corynebacterium pseudotuberculosis causes caseous lymphadenitis (CLA), which is a contagious and chronic disease in sheep and goats. In order to assess the histopathological changes observed in the reproductive organs of nonpregnant does infected with the bacteria, 20 apparently healthy adult Boer does were divided into four inoculation groups, intradermal, intranasal, oral, and control, consisting of five goats each. Excluding the control group, which was unexposed, other does were inoculated with 107 CFU/1 mL of live C. pseudotuberculosis through the various routes stated above. Thirty days after infection, the ovaries, uterus, and iliac lymph nodes were collected for bacterial recovery and molecular detection, as well as histopathological examination. The mean changes in necrosis, congestion, inflammatory cell infiltration, and oedema varied in severity among the ovaries, uterus, and iliac lymph nodes following different inoculation routes. Overall, the intranasal route of inoculation showed more severe (p < 0.05) lesions in all the organs examined. The findings of this study have shown that C. pseudotuberculosis could predispose to infertility resulting from pathological lesions in the uterus and ovaries of does.

Clinico-pathology, hematology, and biochemistry responses toward Pasteurella multocida Type B: 2 via oral and subcutaneous route of infections

Background: Pasteurella multocida a Gram-negative bacterium has been identified as the causative agent of many economically important diseases in a wide range of hosts. Hemorrhagic septicemia is a disease caused by P. multocida serotype B:2 and E:2. The organism causes acute, a highly fatal septicemic disease with high morbidity and mortality in cattle and more susceptible in buffaloes. Therefore, the aim of this study was to investigate the clinical signs, blood parameters, post mortem and histopathology changes caused by P. multocida Type B:2 infections initiated through the oral and subcutaneous routes. Methods: Nine buffalo heifers were divided equally into 3 treatment groups. Group 1 was inoculated orally with 10 ml of phosphate buffer saline; Groups 2 and 3 were inoculated with 10 ml of 1012 colony forming unit of P. multocida Type B:2 subcutaneously and orally respectively. Results: There was a significant difference (p<0.05) in temperature between the subcutaneous and the control group. The results revealed significant differences (p<0.05) in erythrocytes, hemoglobin, packed cell volume, leukocytes, monocytes, and A: G ratio between the subcutaneous and the control group. Furthermore, there were significant differences (p<0.05) in leukocytes, band neutrophils, segmented neutrophils, lymphocytes, eosinophils, basophils, thrombocytes, plasma protein, icterus index, gamma glutamyl tranferase and A: G ratio between the oral and the control group. The post mortem lesions of the subcutaneous group buffaloes showed generalized hyperemia, congestion and hemorrhage of the immune organs, gastro-intestinal tract organs and vital organs. The oral group buffaloes showed mild lesions in the lung and liver. Histologically, there were significant differences (p<0.05) in hemorrhage and congestion; necrosis and degeneration; inflammatory cells infiltration; and edema in between the groups. Conclusion: This study was a proof that oral route infection of P. multocida Type B:2 can be used to stimulate host cell responses where oral vaccine through feed can be developed in the near future.

Assessment of blood changes post-challenge with Corynebacterium pseudotuberculosis and its exotoxin (phospholipase D): A comprehensive study in goat

Aim: There is very little information regarding blood changes during the challenge of phospholipase D (PLD) in goats. Therefore, this experiment was conducted to study the changes in blood after the challenge with Corynebacterium pseudotuberculosis and its exotoxin, PLD to fill in the gap of caseous lymphadenitis (CLA) research. Materials and Methods: Twenty-six crossbred Boer goats aged 12-14 months were divided into 3 groups; the first group n=6 was inoculated with 1 ml phosphate buffered solution s.c. as the control. The second group n=10 was inoculated with C. pseudotuberculosis 1 × 109 cfu s.c. The third group n=10 was intravenous injected with PLD 1 ml/20 kg body weight. Serial blood collections were done at 1 h, 3 h, 5 h, 8 h, and 12 h then every 24 h post-inoculation for the first 30 days of the experiment. Subsequently, the blood collection continued twice a week till the end of the experiment (90 days post-challenge). Results: Both C. pseudotuberculosis and PLD treated groups showed significant changes (p<0.05) in red blood cell count, hemoglobin (Hb), packed cell volume, mean corpuscular volume, mean corpuscular Hb concentration, white blood cell count, neutrophils, lymphocytes, monocytes, eosinophils, basophils, globulin, and total plasma proteins. Similarly, both treated groups showed significant changes (p<0.05) in alanine transaminase, alkaline phosphatase, aspartate transaminase, total bilirubin, calcium concentration, creatine phosphokinase, creatinine, gamma-glutamyl transpeptidase, urea concentration, lactate dehydrogenase, prothrombin time, and activated partial thromboplastin time. Conclusion: It concluded that C. pseudotuberculosis and PLD have a negative impact on the goat’s health in general reflected by all those changes recorded in the hemogram, leukogram, and the blood chemistry.

Clinico-pathological Features in Mice Following Oral Exposure to Pasteurella Multocida B: 2

Haemorrhagic septicaemia (HS) is a major cause of losses to livestock production in many countries around the world. In Malaysia, more specifically, the disease yet remains a major constraint to the national industry. However, the pathogenesis of haemorrhagic septicaemia is another scenario in which the limitations still exists. Thus, the present paper provides more information on the pathogenicity and host response dynamics in a mouse model. Our study of experimental nature manipulates P. multocida serotype B:2, the bacterium responsible for the disease in Asia. In this study, sixteen mice (n=16) were divided into two groups (A & B) of 8 mice each group. Animals in group A were inoculated orally with 1.0 ml 10 9 cfu/ml of P. multocida type B while mice in group 2 were challenged orally with 1.0 ml of phosphate buffer saline (PBS). The animals were observed for clinical signs for 5 days. The mice showing severe signs and surviving mice after 5 days of post- inoculation were euthanized using cervical dislocation approach and the organs such as heart, lung, kidney, stomach, spleen, colon and small intestine were collected for microscopic examinations. The result indicated that mice inoculated with the Pasteurella multocida showed significant (p<0.05) severe clinical signs compared to control group.These clinical signs ranged from mild to severe in which most of individuals infected with Pasteurella multocida showed moderate to severe clinical signs of ruffled hair, laboured breathing, eye discharge and responsiveness with mean levels of 2.13±0.64, 1.88±0.99, 1.50±1.20 and 1.88±0.99 respectively in comparison to the control group. Moreover, mortality rate was recorded between 24 to 50 h post-inoculation in the group that challenged with Pasteurella multocida type B: 2. Microscopically, the extent of visceral tissue damages due to the infection was scored. The interested parameters included pulmonary oedema, presence of inflammatory cells, haemorrhage and necrosis. Of these parameters, animals in infected group showed significant (p<0.05) differences in all most all visceral organs. Lungs, liver and kidney were, in particular, the most predominantly affected tissues. Therefore, oral inoculation of P. multocida type B in mice showed significant clinical response and cellular changes.

Stage II Keratoconjunctivitis in a Goat: A Case report

A two year old Australian Feral male goat weighing 30 kg was presented to the large animal unit of Universiti Putra Malaysia Veterinary Hospital with the clinical signs of conjunctivitis and corneal ulceration with purulent ocular discharge. A sterile swab was taken from the eyefor bacteriological culture and the culture yielded a mixed growth of Pseudomonas aeroginosa and Moraxella caprae.The conditionwas diagnosed as pink eye disease. The goat was administered 3ml of 20mg/kgoxytetracyclinesubconjunctival injection of 0.4ml, intravenous injection of 2.2mg/kg Flunixin meglumine (antipyretic) agent, topical application of terramycine eye ointment and infusion of 2L of 0.6% Nacl solution. The signs of conjunctivitis and corneal ulcersof the eye had substantially reduced five days post treatment. Quarantine of infected animals, good quality feeding and fly control wasrecommendedas preventive measures. Keywords: Pink eye, goat, Moraxella caprae, bacteriology

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.

Isolation and detection of Corynebacterium pseudotuberculosis in the reproductive organs and associated lymph nodes of non-pregnant does experimentally inoculated through intradermal route in chronic form.

Aim: Corynebacterium pseudotuberculosis is the etiological agent of caseous lymphadenitis that affects sheep and goats. This study was designed to determine the presence of the causative organism in the female reproductive organs and associated lymph nodes in non-pregnant does experimentally inoculated through intradermal route in the chronic form. Materials and Methods: 18 non-pregnant healthy Katjang does aged 2-year-old were divided randomly into two groups. The first and second group consists of nine non-pregnant does each and the two groups were subdivided into three subgroups. The first group was experimentally inoculated with 1 ml of 107cfu of live C. pseudotuberculosis through intradermal route, whereas the second group was inoculated with 1 ml phosphate buffer saline (pH 7) solution intradermally. The first group were further subdivided into three subgroups where, the first subgroup (B1) were kept for 30 days post-infection, second subgroup (B2) were kept for 60 days post-infection, and third subgroup (B3) were kept for 90 days. The second group was further subdivided into three subgroups (C1, C2, and C3) where they were kept for 39, 60, and 90 days post-infection, respectively. Results: From this study, there was successful isolation of C. pseudotuberculosis from the reproductive organs of the treatment group after 60 days post-infection. The subgroups (B1, C1, C2, and C3) did not show any presence of the causative organism in the reproductive organs. The second subgroup B2 and third subgroup B3 showed positive isolation of the causative organisms from the ovary, uterine horns, uterus, cervix, vagina, and inguinal lymph node of the experimental non-pregnant does. Conclusion: This study showed that chronic infection of C. pseudotuberculosis via intradermal route may cause effect toward the reproductive organs and may be able to influence the reproductive efficiency of the infected animals.

Haematological, Biochemical And Serum Electrolyte Changes In Non-Pregnant Boer Does Inoculated With Corynebacterium Pseudotuberculosis Via Various Routes

Caseous lymphadenitis is a chronic disease characterized by internal and external abscesses and is caused by Corynebacterium pseudotuberculosis.Thisstudy is designed to measure the hematological, biochemicaland serum electrolyte changes in experimental non-pregnant doesinoculated with Corynebacterium pseudotuberculosis via variousroutes. Little is known about the changes in these parameters through different routes of infection.A total of 20 healthy does (n=20) were divided into 4 groups (intradermal, intranasal, oral and control) of 5 goats each. The three groups were inoculated with 107cfu/1ml of live Corynebacterium pseudotuberculosis, while control is kept unexposed. Following infection, blood samples were collected from the jugular vein for hematological, biochemical and serum electrolyteanalysis. A significantdecrease was observed in RBC count (p<0.05) in the intradermal group, whilenochangesobserved in PCV, Hb, MCV and MCHC parameters. Significant increasein WBC were observed in intradermal, intranasal and oral groups (p<0.05). Slight increase (p<0.05) in monocyte count was observed in intranasal group. A significant reduction (p<0.05) in lymphocytic count was observed for the intranasally inoculated group (p<0.05) and a slight increase (p<0.05) in neutrophil was observed in intranasal and intradermal groups. Biochemically,creatinine and albumin levels increases inintranasal group(p<0.05) and GGT levels were elevated in all the three infected groups (p<0.05).However,there were no significant changes in AST, T.Protein and APT. Serumelectrolyte, revealed a decrease in Calcium (Ca+) concentration in intradermal group with concentration of 2.22mmol/L,intranasal 2.23mmol/L group (p<0.05), and no changes were observed in potassium (K+) and sodium (Na+). The study, therefore, observed increase in WBCs, neutrophils, monocytes, creatinine, GGT, albumin levels and decrease in RBCs, lymphocytes and calcium concentrations on different route of infections.

Knowledge, attitude and perception regarding antimicrobial resistance and usage among ruminant farmers in Selangor, Malaysia

The increasing public health problem of antimicrobial resistance (AMR) has been linked to the extensive antimicrobial use (AMU) in food animals. We conducted a survey among ruminant farmers in Selangor, Malaysia to assess their level of awareness on AMR, attitudes towards AMU, and determinants that influence their practices. The survey was developed in English and Malay, validated, and administered to ruminant farmers in Selangor. A total of 84 farmers (response rate of 55%) completed the structured questionnaire. They appeared to be little aware of AMR and the impact on animals and public health. Indications of inappropriate AMU include their misconception on conditions requiring antibiotic therapy and easy accessibility to antibiotics. More than 70% (60/84) of the respondents believed that all sick animals need to be given antimicrobials. Half of the farmers especially those involved in the production of small and large ruminants; namely mixed ruminant farmers (MRF) (63%, 31/49) indicated that antimicrobials do not have any side effects in animals. Sixty-four percent (54/84) of the farmers have stored antimicrobials in their farms of which the practice was more common (P = 0.02) among the MRF compared to the single ruminant farmers (SRF). Although most of the farmers felt good farm biosecurity will help reduce AMU, they were indifferent regarding using antimicrobials only when prescribed by a veterinarian and non-storage of antimicrobials for later uses. Farmers with larger herd size (>100 animals/herd) and few years of farming experience agreed more to the suggestions about their role and that of veterinarians respectively in reducing the drivers of AMR. These areas might need to be considered by advisors to inform ruminant farmers on AMR and to encourage them for prudent AMU in food-producing animals.

Comparative experimental study of Brucella melitensis and its lipopolysaccharide in mouse model infected via subcutaneous route of exposure

Brucella melitensis is a major zoonotic pathogen in which lipopolysaccharide (LPS) is believed to play a major role in the diseases pathogenesis. To study the immunopathophysiological aspects, we established a mouse model experimentally infected with whole cell of B. melitensis and its lipopolysaccharide via subcutaneous route of exposure. Eighty four mice, BALB/c, both sexes with equal gender distribution and 6-8 weeks-old were randomly assigned into 3 groups. Group 1 (n = 36) were subcutaneoulsy inoculated with 0.4 mL 109 of B. melitensis while group 2 (n = 36) were subcutaneously challenged with 0.4 mL 109 of LPS. Group 3 (n = 12) was challenged subcuatneously with phosphate buffered saline and served as a control group. Animals were observed for clinical signs, haematological and histopathological analysis for a period of 24 days post-inoculation. Our results revealed that B. melitensis infected group demonstrated significant clinical signs and histopathological evidence than LPS infected group. However, both infected groups showed elevated levels of interleukins (IL-1β & IL6), antibody levels (IgM & IgG) as early as 3 days post-infection with predominance in LPS infected group. For hormone analysis, low levels of progesterone, estradiol and testosterone were observed in both B. melitensis and LPS challenged groups throughout the study period. Moreover, in B. melitensis infected groups, the organism was re-isolated from the organs and tissues of gastrointestinal, respiratory and reproductive systems; thereby confirming the possible transmission of the disease dynamics. Moreover, LPS stimulated significantly the innate and acquired immune system without significant systemic dysfunction suggesting the potentiality of the protective properties of this component as an alternative vaccine for brucellosis infection. This report is the first detailed investigation comparing the infection progression and host responses in relation to the immunopathophysiological aspects in mouse model after subcutaneous inoculation with B. melitensis and its lipopolysaccharide.