CATTLE NECROBACTERIOSIS IN THE LIVESTOCK FACILITIES OF THE ALMATY REGION

Necrobacteriosis affects many species of animals. The most susceptible and sensitive to Fusobacterium necrophorum are reindeer, cattle and small cattle, pigs, and rabbits. A constant carriership of the causative agent of necrobacteriosis in the rumen and intestines of ruminants has been established, causative agent is found in food particles during chewing, as well as in feces. The causative agent of necrobacteriosis is widespread in the environment (livestock buildings, walking yards, manure, soil, pastures, stagnant reservoirs, etc.). Infestation of animals occurs when the pathogen enters the injured areas of the skin or mucous membranes of animals. Disturbed blood circulation, cracks and peeling of the horn happen as a result of long-term keeping of animals in damp premices, grazing them in damp, swampy areas, and also maceration of the limb tissues. Four cultures of the causative agent of cattle necrobacteriosis Fusobacterium necrophorum were isolated from sick animals with symptoms of lameness, their biological properties were studied. The pathogenicity of the isolated cultures was studied in laboratory animals. The work was conducted in laboratory and production conditions in "KazSRVI" LLP and at the dairy farm at "Arkabay" human settlement (village) of Talgar district of Almaty region, where stall keeping of animals is practiced. Slices from the diseased hoof of cows were taken at the border of the diseased and healthy tissue. Samples of the selected biological material were plated on Kitt-Tarozzi medium at the sampling site on the farm. The biological material taken from sick animals was studied within several hours after sampling in accordance with the guidelines for laboratory diagnosis of necrobacteriosis. Material for laboratory research (sections from the horny tissue of the hoof on the border with the healthy one) were taken fresh and inoculated on a nutrient medium for anaerobes. The results of cultivation of the necrobacteriosis causative agent on liquid and solid nutrient media under anaerobic conditions are presented. To get rid of the accompanying microflora and obtain a pure culture of F. necrophorum, a bioassay was set on laboratory animals - rabbits. All isolated cultures were highly pathogenic for rabbits. On the 14-15th day after infection, the experimental rabbits died. A pure culture of F. necrophorum, not contaminated with extraneous microflora, was sown from the internal organs of rabbits. It was found that rabbits are the optimal biomodel for purification of the F. necrophorum culture. The biochemical properties of the isolated cultures have been studied. It was found that epizootic cultures of the causative agent of necrobacteriosis emitted hydrogen sulfide and had hemolytic properties. In experiments in vitro and in vivo, it was found that the isolated cultures of F. necrophorum showed hyaluronidase activity. Cultures of F. necrophorum had a high catalase activity, they split hydrogen peroxide with the formation of oxygen (gas bubbles). When studying biochemical properties, it was found that F. necrophorum releases ammonia within 2-3 hours. Four cultures of F. necrophorum isolated from biological material from cattle were identical in biological properties.

Lemierre’s syndrome in a child

Lemierre’s Syndrome (LS) is a rare syndrome most frequently due to an anaerobic organism, Fusobacterium Necrophorum. It is commonly a complication of an acute oropharyngeal infection, but there are exceptions to its presentations. In our case the cause of LS was otitis media caused by Streptococcus species. This is a rather unusual presentation of LS. LS is caused due to septic complications of oropharyngeal infections, which lead to thrombophlebitis of internal jugular vein leading to thrombosis formation. In this case report, we present a case of Lemierre’s syndrome in a seven-year-old male child. The patient presented with high grade fever spikes and earache, which were unresponsive to oral antibiotics. LS was diagnosed in this patient on the basis of clinical, microbiological and radiological findings. After the diagnosis, treatment involved using broad spectrum antibiotics and anticoagulants, followed by surgery. Though role of anticoagulants is controversial in LS, but there is no specific guideline contraindicating the use of anti-coagulants. In our case, timely diagnosis and management enabled us to discharge the patient without any symptoms. doi: https://doi.org/10.12669/pjms.38.ICON-2022.5773 How to cite this:Hemani F, Naveed A, Akhtar S, Shahid S. Lemierre’s syndrome in a child. Pak J Med Sci. 2022;38(2):433-435. doi: https://doi.org/10.12669/pjms.38.ICON-2022.5773 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Evaluation of Immunoprotective Effects of Fusobacterium necrophorum Outer Membrane Proteins 43K OMP, Leukotoxin and Hemolysin Multi-Component Recombinant Subunit Vaccine in Mice

We evaluated the efficacy of three vaccine formulations containing different combinations of proteins (43K OMP, leukotoxin recombinant protein PL4 and hemolysin recombinant protein H2) and killed whole cell Fusobacterium necrophorum in preventing liver abscess. Four subcutaneous vaccines were formulated: vaccine 1 (43K OMP), vaccine 2 (PL4 and H2), vaccine 3 (43K OMP, PL4 and H2), and vaccine 4 (killed whole bacterial cell). 43K OMP, PL4, and H2 proteins were produced by using recombinant protein expression. To evaluate vaccine efficacy, we randomly allocated 50 BALB/c female mice to one of five different treatment groups: PBS control group, vaccine 1, vaccine 2, vaccine 3, and vaccine 4. Mice were vaccinated three times, with 14 days between each immunization. After immunization, the mice were challenged with F. necrophorum. The three key findings of this study are as follows: (1) Vaccine 3 has enabled mice to produce higher antibody titer following bacterial challenge, (2) in the liver pathology of mice, the vaccine 3 liver showed the least pathology, and (3) all four vaccines produced high levels of antibodies and cytokines in mice, but the level of vaccine 3 was the highest. Based on our results, it has been demonstrated that a mixture of F. necrophorum 43K OMP, PL4, and H2 proteins inoculated with mice can achieve protection against liver abscess in mice. Our research may therefore provide the basis for the development of a vaccine against F. necrophorum bovine infections.

Impacts of polyclonal antibody preparations from avian origin as a feed additive to beef cattle: Immune responses during the step-up transition diets

This study investigated the effects of feeding an avian-derived polyclonal antibody preparation (PAP; CAMAS, Inc.) against Streptococcus bovis, Fusobacterium necrophorum, and lipopolysaccharides (40, 35, and 25% of the preparation, respectively) on immune responses [haptoglobin (Hp), serum amyloid A (SAA), rectal temperature (RT), leukocyte counts, and expression of cell adhesion molecules cluster of differentiation (CD) CD11b, CD14, and CD62L] of beef steers during a 21-d step-up adaptation to a high-grain diet. Eight ruminally cannulated Angus crossbred beef steers (658 ± 79 kg of BW) were assigned in a cross-over design and transitioned from a diet containing bermudagrass hay [Cynodon dactylon (L.) Pers.] ad libitum plus 0.45 kg/d of molasses with 0 (CON) or 3 g of PAP (PAP) to a high-grain diet. Transition consisted of three 7-d steps of increased inclusion of cracked corn (35, 60, and 82% of the diet dry matter for STEP1, STEP2, and STEP3, respectively). On each transition day and 7 d after STEP3 (STEP3-7d), RT was obtained every 3 h for a total of 24 h, whereas blood was collected on d 0, 1, and 3, relative to diet transition. There were no effects of PAP inclusion in any of the blood parameters (P > 0.11). However, a tendency for day effect (P = 0.10) was observed for concentrations of Hp, which were greater on d 3 and 7 vs. d 0 relative to the second diet transition (STEP2). Plasma concentrations of SAA were greater on d 1, 3, and 7 compared to d 0 during STEP1 (P = 0.01), while during STEP2 and STEP3, SAA concentrations increased (P < 0.01) from d 0 to 3. During STEP2, PAP steers tended to have lower (P = 0.08) RT than CON steers. Neutrophil and monocyte counts were the least during STEP3 (P < 0.01), whereas expression of CD11b and CD62L was the least through forage feeding (P < 0.01). Concentration of starch in the diet was correlated to all the variables tested (P ≤ 0.01), except for the percentage of B cells (P = 0.22). Yet only ruminal pH, RT, monocyte, and neutrophil counts presented strong correlation coefficients. In conclusion, the step-up transition from forage to high-grain diets triggered systemic inflammation in beef steers as observed by increased plasma concentrations of haptoglobin, serum amyloid A, and expression on adhesion molecules in leukocytes. However, feeding polyclonal antibody preparations against Streptococcus bovis, Fusobacterium necrophorum, and lipopolysaccharide did not provide benefits to mitigate inflammation.

Evaluating the Efficiency of Newly Formulated Pomade® and Ceftiofur Hydrochloride for Treating Foot Rot in Dairy Cattle

Background: Foot rot is an important contagious disease that causes economic loss in dairy cattle. Even though many antibiotic treatments have been tried on foot rot, very few information about new topical treatment method or product have been reported about the disease. The main objective of this paper is to evaluate an alternative new treatment for foot root disease in dairy cattle. Methods: Forty-one swap samples were collected from cattle’s feet which were further investigated under microscopic examination and polymerase chain reaction (PCR) for Dichelobacter nodosus and Fusobacterium necrophorum. Newly formulated pomade® and ceftiofur hydrochloride (Eficur®) applications were used in foot rot cases. Result: Polymerized chain reaction revealed D. nodosus in all the samples were as F. necrophorum was seen only in 22 (53.66%) samples. In this study, D. nodosus was considered as the primary agent involved in foot disease of cattle and F. necrophorum along with other bacterias were considered to be associated in the infection. The foot rot wounds formed in all cases (100%) in which D. nodosus, F. necrophorum and other bacteria were detected were healed along with tissue regeneration. As a result of treatment applications, a success rate of 93.33% was obtained in lameness resulting due to medium foot rot cases and 45.45% in severe acute lameness. The lameness recovery rate was found to be 80.48%.