Hematological Variations Associated with Bovine Foot and Mouth Disease Virus Infection

Transcription

1 Journal of Veterinary Advances Hematological Variations Associated with Bovine Foot and Mouth Disease Virus Infection Olabode H. O. K., Kazeem H. M., Raji M. A., Ibrahim N. D. G., Adah B. M. J. and Obafemi F. A. J Vet Adv 2013, 3(9): DOI: /jva Online version is available on:

2 ISSN: OLABODE ET AL. Original Article Hematological Variations Associated with Bovine Foot and Mouth Disease Virus Infection 1,2 Olabode H. O. K., 2 Kazeem H. M., 2 Raji M. A., 2 Ibrahim N. D. G., 1 Adah B. M. J. and 3 Obafemi F. A. 1 Department of Veterinary Microbiology, Faculty of Veterinary Medicine, University of Abuja. 2 Department of Veterinary Microbiology, Faculty of Veterinary Medicine, Ahmadu Bello University, Zaria. 3 Department of Veterinary Physiology and Biochemistry, Faculty of Veterinary Medicine, University of Abuja. Abstract Hematological analysis of bovine blood was carried out to establish possible variations associated with foot and mouth disease (FMD) infection. Random sampling of FMD infected bovine blood from the reported outbreaks in Bode, Bode II and Buakaru-rotuwa wards in ilesha baruba, Baruten LGA in kwara state was collected forth nightly in December, 2010, January, February, and March, Serum antibodies detection using NS-ELISA revealed out of the ninety cattle sampled in the fifteen outbreaks, 64 showed positive evidence of FMD with percentage distribution of 37.5% and 62.5% for bulls and cows respectively. This was significant (p<0.05) by chi square analysis. The overall seropositivity of 71.11% recorded showed breed seropositivity of 64.9% and 75.5% for bulls and cows respectively. The clinical hematological picture of FMD outbreak seropositive reactors revealed anemia, leucopenia and leucocytosis, as distinct changes observed. The anemia observed was characterized by anisocytosis and poikilocytosis. However, this characteristic features of the blood showed no significant (P>0.05) association with FMD sero-positive cattle by chi square analysis. Therefore, this finding provides preliminary information on hematological variation observed during FMD outbreak. Further experimental case-control study on the hematology of FMD infected cattle is thus suggested. Keywords: Hematology, bovine, variation, foot and mouth disease, outbreaks. Corresponding author: Department of Veterinary Microbiology, Faculty of Veterinary Medicine, University of Abuja, Ahmadu Bello University, Zaria. Received on: 13 Jun 2013 Revised on: 10 Jul 2013 Accepted on: 25 Sep 2013 Online Published on: 30 Sep J. Vet. Adv., 2013, 3(9):

3 Introduction Foot and mouth disease (FMD) also known as aphthous fever or infectious aphthous stomatitis (Kitching, 2002a) is a highly contagious viral transboundary disease of both domestic and wild cloven hoofed animals (Di Nardo et al., 2011). This disease has constraint international trade of animal products and FMD control to protect the livestock industries of industrialized countries as well as the livelihoods and income generation of developing countries where FMD continues to be endemic (Rweyemamu et al., 2008) especially in Nigeria (Abdulkadir, 1989; Chukwuedo et al., 2008) with serious impact on social and economic consequences of reduced milk and meat production as result of high morbidity and loss of market value (Sangare et al., 2004). In Africa, cattle and buffalo (Syncerus caffer) are usually the common host of FMD viruses. Of the domesticated species, cattle, pigs, sheep, goats and buffalo are most susceptible, also many species of cloven-hoofed wildlife, such as deer, antelope and wild pigs may become infected (FAO, 1984). The disease in cattle is usually obvious in the unvaccinated herds. However, in vaccinated herds and in some breeds indigenous to areas in which FMD is endemic, it may circulate undetected (Kitching, 2002b). The classical clinical features of FMD in cloven hoofed mammals occur following 2-14 days Incubation period as described (Remond et al., 2002). Initial sign includes, pyrexia [up to 106 F (40oC) lasting 1-2 days before other clinical signs], pyrexia is followed by anorexia, agalactia in milking animals and appearance of vesicles (Clavijo et al., 2004). Painful lesions cause profuse drooling, bruxism, foot stamping, and lip smacking with rupture of vesicles occurring within hours to 2 days, leaving erosions and recovery generally occurs within 8-15 days. FMD virus (Asia 1) was reported to have caused mild type disease which induced transient moderate leucopenia with lymphopenia in experimental cattle and buffaloes but with diverse levels of monocytes. This microscopic change was however, observed in keratinized epithelium of tongue and foot (Mohan et al., 2008). However, there is paucity of reports on hematologic features associated with FMD in the HEMATOLOGICAL VARIATIONS ASSOCIATED WITH... study area. This premised the search for associated changes in blood picture of bovine FMD outbreaks in affected herds under field condition, in order to avail clinicians, researchers and field practitioners with additional information in the understanding of the patho-physiology of FMD. Materials and Methods Study Area Kwara State is also referred to as the gateway State because of its location between the Northern and Southern parts of Nigeria. This State is a first order administrative division located on the longitude N 8 30' 0'' and latitude E 5 0' 0'' 8.5 / 5 (Geo Name Id : ) (Anon, 2010) with 16 Local Government Areas namely: Asa, Baruten, Edu, Ekiti, Ifelodun, Ilorin East, Ilorin West, Irepodun, Isin, Kaiama, Moro, Offa, Oke-Ero, Oyun, Ilorin south and Pategi. This state shares common boundaries with Niger and Kebbi States to the North, Oyo, Ondo and Edo States to the South, Benue, Plateau and Federal Capital Territory to the East (2011). It maintains an international boundary with the Republic of Benin to the West. Study Design A random sampling of FMD infected cattle based on the evidence of recent outbreak report, presence of rupture and unruptured vesicles in the buccal and or around the feet as well as the willingness of the cattle owners at various location covered by this study. The sampling was conducted forth nightly over a period of three months (December, 2010, January, February, and March, 2011) within 3-10days post outbreak report. Three locations namely Bode, Bode II and Buakarurotuwa wards in ilesha baruba, Baruten, LGA in kwara state was considered based on their locations and proximity to cattle trade routes and market. Ninety (90) blood samples were collected from the fifteen (15) reported outbreaks in herds with no history of FMD vaccination. Sample Collection Whole blood was also collected using 10mls Syringe and 18G 11/2 needle through the Jugular vein of well restrained FMD affected cattle during outbreak investigation. Halve (5ml) was transferred 246 J. Vet. Adv., 2013, 3(9):

4 OLABODE ET AL. into 5ml EDTA containing plastic Container bottles (maxicomr) for whole blood, while the other halve (5ml) was transferred into 5ml plain plastic container (ANTECR) for sera collection. All the samples were packed on ice and transported to the laboratory, National Veterinary Research Institute, Vom and stored until use at 4oC and -70oC respectively. Serological Assay Using FMDV-NS ELISA FMDV NS for in vitro detection of antibodies against bovine FMDV in sera samples as described by Sorensen et al., (1998) was conducted using PrioCHECKR FMDV-NS blocking ELISA. The preparation of working solutions was conducted in advance, this include Dilution buffer working solution, Additives, ELISA buffer, Conjugate dilution and Washing solution as recommended by the manufactures [Prionics Lelystad B.V. :The Netherlands]. Then percentage Inhibition (PI) was calculated and interpreted as PI of < 50% was considered negative and it was interpreted that the animal tested had not been exposed to FMD for 40 days. PI of 50% was considered positive and recent exposure <40> days to FMD. More specifically, PI value of 50% but < 70% was considered a weak positive result and PI value of 70% was considered a strong positive result (Sorensen et al., 1998). Hematological Analysis The corresponding pair of the whole blood collected during FMD outbreaks in Ilesha baruba was be subjected to hematological techniques as described (Jain, 1993; Kumshe et al., 1997). The Packed Cell Volume (PVC) was determined by the microhaematrocit technique and total white blood cell counts were done by the use of Neubauer haemocytometer. The values were noted for association with FMD. Statistical Analysis The positive reactors were expressed as simple descriptive statistics such as percentage and both positive and negative FMD reactors were compared by Chi square. The hematology data was also subjected to Chi square analysis using SPSS 19 software. Results Detection of serum antibodies using NS-ELISA revealed out of the ninety cattle sampled in the fifteen outbreaks, 64 showed positive evidence of FMD with percentage distribution of 37.5% and 62.5% for bulls and cows respectively. This was significant (p<0.05) by Chi square analysis with X2=53.0 df= 1. The overall seropositivity of 71.11% recorded showed breed seropositivity of 64.9% and 75.5% for bulls and cows respectively as indicated in Table 1. The clinical hematological picture of FMD outbreak sero-positive reactors in Ilesha Baruba as shown in Table 2 revealed that 71.0% (27/38) were anemic, 70% (21/30) showed leucopenia, 86% (6/7) indicated leucocytosis, 2 showed anisocytosis while 3 presented poikilocytosis and 60% (7/10) showed both anisocytosis and poikilocytosis. However, this characteristic features of the blood showed no significant (P>0.05) association with FMD seropositive cattle by Chi square analysis with X2 value of 5.0, df=5 with some expected frequency less than 1. Table 1: Showing FMD sero-positive reactors in outbreak in Ilesha Baruba. Sex No. Positive No. Negative Total Percentage % Bulls Cows Total J. Vet. Adv., 2013, 3(9):

5 HEMATOLOGICAL VARIATIONS ASSOCIATED WITH... Table 2: Hematological picture of FMD outbreak sero-positive reactors in Ilesha Baruba. Blood parameter FMD sero-positive FMD sero-negative Total reactors (%) reactors (%) Anemia (PCV< 24) 27 (71.1%) 11 (28.9%) 38 Leucopenia (TWBC < 21 (70.0%) 9 (30.0%) x10 9 L) Leucocyctosis 6 (85.7%) 1 (14.3%) 7 (TWBC > 12.0x10 9 L) Anisocytosis 2 (100%) - 2 Poikilocytosis 3 (100%) - 3 Both Anisocytosis Poikilocytosis 7 (70%) 3 (30%) 10 Discussion FMD outbreak examination by most clinicians in Nigeria is largely based on the presences of ruptured or unruptured vesicles (clinical signs and herd history) with little emphasis on molecular characterization of FMDV isolates due to insufficient field and laboratory facilities which necessitated the search for alternative simple hematologic indicators that may aid identification of FMD affected cattle. However, antibodies to FMD viral nonstructural proteins (NSPs) have often been used as indicators of infection, irrespective of vaccination status (OIE, 2008). Detection of serum antibodies using NS-ELISA in this study revealed out of the ninety cattle sampled within 3-10days post infection from the fifteen reported outbreaks indicated that 37 were bulls and 53 cows were sampled with an overall FMD sero-positivity 64 (71.11%) as indicated in Table 1. This conforms to report that affirm cattle exposed to FMDV within minimum <40> days can be detected using NS ELISA (Sorensen et al., 1998). The occurrence of sero-positive reactors in these outbreaks connotes other previous documented outbreak reports around the country (Durojaiye, 1981; Abegunde, 1987; Knowles et al., 2008). The clinical hematological picture of FMD outbreak sero-positive reactors as shown in Table 2 revealed anemia, leucopenia and leucocytosis, as distinct changes observed in this study. The anemia observed was by characterized anisocytosis and poikilocytosis. This connotes earlier report (Radostitis et al., 2000) that anemia occurred in bovine FMD but as a sequel probably due to endocrine damage which lead to chronic syndrome. In addition, hemoparasitism and immunosupression that may arise from other management stress could also be incriminated as a contributing factor to the observed anemia; this was not clarified as that was not in the scope of this study. The leucopenia observed in this study, connotes previous reports of Reece, (2009) that leucopenia was usually associated with early stages of viral infections as the blood samples were collected within 3-10days post outbreak reports as well as that reported by Mohan et al., (2008) in cattle and buffaloes infected with FMD virus (Asia 1). Leucocytosis is an increase in leukocyte number which usually occurs in bacterial infection (Reece, 2009) was also observed in some FMD positive reactors. This could be associated with ongoing bacterial infections. Although plasma biochemical parameters was not analyzed, these characteristic features of the blood showed no significant (P>0.05) association with FMD sero-positive cattle by Chi square analysis. In conclusion, the finding of this study provides preliminary information on the hematological characteristics observed in FMD affected cattle under field exposure condition. Further experimental case-control study on the hematology of FMD infected cattle is thus suggested. Acknowledgement Authors wishes to appreciate the efforts Mr. Andrew Onoja and Mrs. Deborah of the clinical pathology, Central diagnostic laboratory, for assisting in the hematological analysis. Also Dr. Ularamu, H.G, Dr. Wugak, Y.S and Mr. Agom of the FMD laboratory National Veterinary Research 248 J. Vet. Adv., 2013, 3(9):

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