1 1 Research article 2 Title: 3 Dairy cattle mortality in an organized farm in Bangladesh 4 5 Authors: 6 M. M. Hossain1, 3*, A. H. M. Kamal1, A. K. M. A. Rahman2, H. S. Cho3 7 8 1 Department of Medicine, Sylhet Agricultural University, Bangladesh 9 2 Department of Medicine, Bangladesh Agricultural University, Mymensingh 10 11 12 3 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 College of Veterinary Medicine and Bio-safety Research Institute, Chonbuk National University, Jeonju 561-756, Republic of Korea *Corresponding Author: Md. Mukter Hossain DVM, MS, MSc PhD Student College of Veterinary Medicine Chonbuk National University Jeonju 561-756, South Korea Phone: +82-63-270-4872 (Office) Cell:+82-10-8200-5510 Fax:+82-63-270-3780 E-mail: mukter.sau@gmail.com 33 1 2 34 Dairy cattle mortality in an organized farm in Bangladesh 35 36 ABSTRACT 37 A retrospective study of dairy cattle mortality on the Central Cattle Breeding and Dairy Farm in 38 Bangladesh was carried out between 1992 and 2007. Sixteen years of data on mortality of dairy 39 cattle were analyzed for the effects of year, season, age, sex, breed, and etiology on mortality 40 rate. The average overall mortality rate was 5.60% and on average, female cattle (55.71%) were 41 found to die more than males (44.29%). Mortality was significantly higher in exotic and 42 crossbred animals than in local/indigenous cattle. There was variation in seasonal mortality rate 43 averaging 28.99%, 33.03%, and 37.98% in rainy, summer, and winter seasons, respectively. The 44 major causes of death were diseases of the respiratory tract, mainly pneumonia (39.91%). 45 Tuberculosis was the second most common cause of mortality accounting for 20.58% of deaths. 46 The other major cause of death was disease of the alimentary tract, mainly enteritis (15.58%). 47 Other causes of death occurred in the following frequencies: malnutrition (5.91%), debility 48 (4.43%), hairball (3.35%), tympanitis (2.56%), babesiosis (2.27%), internal hemorrhage (2.16%), 49 black quarter (1.76%), and foot and mouth disease (1.48%). Of the four potential risk factors 50 investigated, age was the most important factor and significantly associated with mortality. 51 During the first month of life, calves had a higher risk of mortality than adults. 52 53 Key words: Mortality, Cattle, Causes, CCBDF, Bangladesh 54 55 Introduction 56 Dairy cattle mortality is a severe problem for the dairy industry. Mortality among dairy cattle 57 results in financial loss, including the value of the lost cattle, cost of replacement, loss of milk 2 3 58 production, and extra labor (Thomsen et al., 2006). Mortality patterns in organized dairy herds 59 serve as a useful indicator for assessing the status of herd health and the efficacy of management 60 programs (Prasad et al., 2004). A rise in mortality among a group of cattle can indicate sub- 61 optimal health and welfare (Thomsen et al., 2004). Bangladesh is a densely populated country 62 with an agricultural-based economy and its livestock play an important role. Agricultural 63 livestock account for about 6.5% of total income and about 25% of the country’s population are 64 directly involved in this sector. Dairy industry is a crucial component of an agro-based economy 65 for a country like Bangladesh (Islam et al., 2006). Inadequate feeding, widespread disease, and 66 inefficient extension services are considered among the most binding constraints on livestock 67 development in Bangladesh. 68 69 The Directorate of Livestock Services (DLS), Government of Bangladesh has established a 70 Central Cattle Breeding and Dairy Farm (CCBDF) at Savar, Dhaka to meet the excessive 71 demands for milk, particularly for the capital city. The success of the CCBDF partly depends on 72 sound health management of the animals and efficient production techniques. However, diseases 73 are one of the major health constraints for livestock development in Bangladesh. Calf mortality 74 in every dairy and breeding farm results in financial and genetic loss. Cattle mortality was shown 75 to be the major cause of economic losses on a dairy farm in Bangladesh (Islam et al., 2005). 76 Therefore, reductions in morbidity and mortality rate are the first and foremost targets of dairy 77 farm management (Prasad et al., 2004). Identification of factors that are responsible for the death 78 of cattle is an important prerequisite for avoiding excessive mortality. An epidemiological 79 approach usually involves collecting and evaluating pre-existing data to find associations with 80 the problem. 3 4 81 MATERIALS AND METHODS 82 Study site: The study was conducted at the CCBDF at Savar, Dhaka, Bangladesh. The CCBDF 83 was established in 1973 on 1300 acres of land with the assistance of the German Agency for 84 Technical Cooperation at Savar Upazila in the Dhaka district of Bangladesh. This farm is located 85 between 23O 46’ and 23 O 58’ North latitude and 90 O 12’ and 90 O 20’ East longitude and about 86 30 km northwest of the capital city of Dhaka. 87 Nature and source of data: For the retrospective study, the data were collected based on farm 88 records of all cattle deaths over a period of 16 years (1992 to 2007) on the CCBDF. Information 89 on the date of birth, sex, breed, date of death, and causes of death were collected from the farm at 90 the individual animal level. The age and breed of cattle were collected from records available in 91 the byre section of the farm. Initially all data were recorded on a day-to-day basis and were then 92 tabulated on a master sheet on a week-by week and month-by-month basis to assess weekly, 93 monthly, and yearly variations. 94 Data on causes of mortality: Data on the causes of mortality were obtained from death reports 95 routinely prepared by veterinarians at the Veterinary Hospital of the CCBDF. Each death report 96 usually consisted of history, clinical and macroscopic post-mortem findings, and occasionally 97 laboratory data. However, because most diagnoses were based on clinical grounds and post- 98 mortem findings and in such cases confirmed by laboratory aids, the data were arranged into five 99 major diagnostic groups as follows: (1) bacterial and mycobacterial diseases (black quarter and 100 tuberculosis); (2) viral disease (foot and mouth disease); (3) protozoan disease (babesiosis); (4) 101 disease syndrome (debility, enteritis, internal hemorrhage, malnutrition, pneumonia, and 102 tympanitis); and (5) physiological disorder (hair ball). The data were checked manually for 103 obvious inconsistencies, recording errors, or missing data. 4 5 104 Statistical analysis: Microsoft Excel 2003 was used to calculate the descriptive statistics. Then 105 the collected data were transferred to STATA version 11 (Stata Corporation, Texas, USA, 2009) 106 for statistical analysis. The Poisson regression model was used to analyze the mortality of dairy 107 cattle. All tests of statistical significance were carried out at the p <0.05 level unless otherwise 108 stated. 109 Results 110 Mortality: A total 1759 animals died during the 16 years (1992–2007) that were evaluated. 111 Among them, female cattle died more than males and the average overall mortality rate was 112 5.60%. The estimated annual mortality rates and pattern of dairy cattle are illustrated in Figure 1. 113 Trends in mortality indicated that the highest number of dairy cattle died in 2004 which 114 accounted for 215 (11.58%) deaths. This was approximately twice the number of deaths 115 compared with the previous 12 years. There was a lower level of cattle mortality rate (CMR) for 116 the period of 1992 to 1999 (in those 8 years CMR varied between 1.05% and 6.07%) and a 117 higher rate for the period of 2000 to 2007 during which CMR varied between 1.16% and 118 11.58%. 5 6 14 12 10 % Mortality 8 6 4 2 0 119 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 Year 120 121 Figure 1. Annual mortality rates by year for dairy cattle on the CCBDF, Savar, Bangladesh 122 (1992–2007). 123 124 Factors associated with mortality: 125 Age and Sex: Mortality among dairy cattle was highest up to 365 days of age and in female 126 calves, which accounted for 40.31% of deaths. The lowest mortality among dairy cattle was 127 estimated in heifers (5.06%). 128 Table 1. Descriptive statistics and Poisson regression analysis by age of dairy cattle mortality on 129 the CCBDF, Savar, Bangladesh (1992–2007). 130 Number P-value a Category of animal 95% confidence interval a of deaths Female calves ≤ 365 days 709 0.000 1.968032 2.621291 6 7 Male calves ≤ 365 days 540 0.000 1.184095 1.559081 Heifers 365 days to ≤ 730 days 89 0.258 -.1605987 .5994834 102 0.002 -.7171178 -.1634381 Bulls ≥ 731 days 137 0.037 .0324649 1.062232 Cows ≥ 731 days 182 0.000 .6494502 1.34968 Young bulls 365 days to ≤ 730 days 131 a Used in Poisson regression. 132 133 Calves (71.01%) had higher mortality compared with young (10.85%) and adult cattle (18.13%). 134 These differences are statistically significant (Table 1). Higher percentage of deaths was 135 observed within 365 days of age and the mortality started to decrease gradually after their first 136 month of life and dropped to a lower rate with the increase of age. This low rate of mortality was 137 maintained during the remaining part of one year of age with slight variations between different 138 age intervals (Table 2). 139 140 Table 2. Relationship of calf mortality of different intervals age in CCBDF, Savar, Bangladesh 141 (1992-2007) No. of deaths Mortality (%) Age (days) Female Male Total Female Male Total 1-30 204 168 372 28.77 31.11 59.88 31-60 157 118 275 22.14 21.85 43.99 7 8 61-90 101 68 169 14.25 12.59 26.84 91-120 56 38 94 7.90 7.04 14.94 121-150 34 30 64 4.80 5.56 10.36 151-180 21 17 38 2.96 3.15 6.11 181-210 15 14 29 2.12 2.59 4.71 211-240 27 11 38 3.81 2.04 5.85 241-270 22 16 38 3.10 2.96 6.06 271-300 21 15 36 2.96 2.78 5.74 301-330 21 18 39 2.96 3.33 6.29 331-365 30 27 57 4.23 5.00 9.23 142 143 Breed and Season: The highest number of cattle deaths was in the Local × Friesian herd 144 category (52.70%) followed by Sindhi × Friesian (15.97), Australian × Friesian × Sahiwal (8.64), 145 Local × Sindhi × Friesian (8.07), Local (4.49), Sindhi (4.32), Friesian (3.13), and 2.67% cases of 146 Australian x Friesian × Sahiwal × Friesian. Table 3 depicts the seasonal mortality rates among 147 different breeds. The seasonal comparison of mortality rates indicated that the overall mortality 148 rate varied between 28.99% (rainy season) to 37.98% (winter season). 149 150 Table 3. Seasonal mortality (%) in different breeds of dairy cattlea on the CCBDF, Savar, 151 Bangladesh (1992–2007). Season Season AFS AFSF F L LF LSIF SIF SI total 8 9 Rainy 8.30 1.88 2.56 3.87 11.43 0.63 0.06 0.28 28.99 (146)b (33) (45) (68) (201) (11) (1) (5) (510) 0.06 0.40 0.17 0.34 31.67 0.17 0.11 0.11 33.03 (1) (7) (3) (6) (557) (3) (2) (2) (581) 0.28 0.40 0.40 0.28 9.61 7.28 15.80 3.92 37.98 (5) (7) (7) (5) (169) (128) (278) (69) (668) 8.64 2.67 3.13 4.49 52.70 8.07 15.97 4.32 100.00 (152) (47) (55) (79) (927) (142) (281) (76) (1759) Summer Winter Overall 152 a 153 = Holstein Friesian; L = Local; LF = Local x Friesian; LSIF = Local × Sindhi × Friesian; SIF = 154 Sindhi × Friesian; and SI = Sindhi. 155 b AFS = Australian × Friesian × Sahiwal; AFSF = Australian × Friesian × Sahiwal × Friesian; F Values in parentheses indicate the total numbers of cattle that died during each season. 156 157 Causes of mortality: A total of 11 types of diseases and disease syndromes responsible for the 158 death of dairy cattle were identified at the CCBDF Veterinary Hospital. The overall mortality 159 rates per disease or disease syndrome is shown in Table 4. 160 9 10 161 Table 4. Specific diagnosis of diseases & syndromes responsible for dairy cattle mortality on the 162 CCBDF, Savar, Bangladesh (1992-2007) Proportional mortality rate Diseases & syndrome No. of deaths (n=1759 dead) Black quarter 31 1.76 Tuberculosis 362 20.58 FMD 26 1.48 Babesiosis 40 2.27 Debility 78 4.43 Enteritis 274 15.58 Internal Haemorrhage 38 2.16 Malnutrition 104 5.91 Pneumonia 702 39.91 Tympanitis 45 2.56 Hairball 59 3.35 163 164 A total of 393 (22.34%) deaths were estimated to be from bacterial and mycobacterium diseases 165 in which tuberculosis (20.58%) and BQ (1.76%). Of viral diseases, 26 (1.8%) of total cases were 166 associated with FMD whereby it is higher in female animals (84.62%). With regards to protozoal 167 diseases, total of 40 (2.27%) death were from babesiosis and mainly in female animals (97.50%) 168 compared to males (2.50%).Calves had the greatest mortality accounting for 80.0% of deaths. 169 Other disease syndrome accounts 55.49% of all deaths, where by pneumonia (39.91%) and 170 enteritis (15.58%). Mortality caused by enteritis and pneumonia also showed makeable 171 associations to the age and sex of the animal. Of all deaths, 197 (11.20%) female calves (< 365 172 days) died from enteritis, and 238 (13.53%) female and 246 (13.99%) male calves were 173 associated with pneumonia. A higher percentage of calves (74.36%) died from debility. 10 11 174 Malnutrition accounted for 5.91% of the deaths, and 45 (2.56%) cases of tympani were 175 diagnosed by history and post-mortem findings. Physiological disorder (hairball) contributed to 176 3.35% of the overall mortality rate, with the highest in female calves (64.41%) followed by cows 177 (25.42%), heifers (6.78%), young bulls (1.69%), and male calves (1.69%). 178 179 Discussion 180 Factors associated with mortality: 181 Age and Sex: Among the deaths, more than two-thirds of mortality occurred within the first year 182 of life with a greater percentage occurring in the first months. These findings are similar to 183 previously reported findings (Debnath et al., 1995; Islam et al., 2005; Martin et al., 1975). 184 Young calves are highly susceptible to infectious diseases as a result of lower immunity which 185 can be exacerbated by in adequate colostrum intake ( Brenner et al., 1989; Oluokun et al., 1988). 186 Although calf mortality on smallholder traditional dairy farms in Bangladesh has been estimated 187 to be relatively lower (Debnath et al., 1990), this estimate is not applicable to the CCBDF since 188 it follows an intensive animal husbandry system which differs markedly from the traditional 189 extensive husbandry systems in villages. Female cattle had higher mortality rates than males, 190 possibly due to the presence of a higher proportion of female animals on the farm. This is likely 191 to reflect their relative value; females give birth to the calves which then join the milking herd 192 and provide a regular income. Similar findings were also reported in a study of mortality pattern 193 in a closed herd of dairy cattle in India ( Somavanshi, 1995 ). 194 195 Breed and Season: The rate of mortality was higher in crossbred cattle compared with 196 indigenous/local and other breeds available on the CCBDF. The higher mortality rate in 197 crossbred cattle may be explained by the prevailing environmental conditions, including 11 12 198 management and climate. Higher mortality in exotic and crossbred cattle could be expected in 199 tropical climates because of greater susceptibility to climatic and disease stresses in such 200 environments. The high rate of mortality in crossbred (Local × Holstein Friesian) cattle may 201 indicate lower survivability of this breed compared with other breeds under the prevailing 202 conditions of Bangladesh. This observation is in agreement with previous findings of higher 203 mortality in exotic and crossbred cattle (Debnath et al., 1995). The largest number of cattle died 204 in the winter season followed by the summer and rainy seasons. Higher mortality in crossbred 205 cattle was observed during the summer season compared with local/indigenous and other sub- 206 continent breeds. The trends of this study indicated that local/indigenous breeds had better 207 adaptability for hot humid weather (summer) and higher susceptibility to cold (winter), whereas 208 the opposite trend was seen in crossbreds. 209 210 Causes of mortality: Tuberculosis was the second most important cause of death in dairy cattle 211 and mortality was more common in calves and during the winter season. These findings are in 212 contrast with previous studies (Mitra et al., 2005; Ramesh, 2007). In a confined herd like on the 213 CCBDF, tuberculosis can be controlled and eradicated by following a “detect and discard” 214 policy. Culling to reduce the population density can also decrease transmission of the disease. 215 The death of cattle by BQ was lower compared with other diseases, possibly due to treatment of 216 infected animals and vaccination against the disease. Among the deaths by BQ, a higher 217 proportion was observed in calves within first year of life. Similarly, Sultana et al. (2008) 218 reported that animals up to 12 months of age have a 6.66 times higher probability of mortality 219 than animals over 12 months of age. Death by FMD showed marked age-specificity, with calves 220 having the highest proportion of deaths. It should be pointed out that in a previous study; FMD 12 13 221 was shown to contribute significantly to calf mortality even after regular vaccination (Debnath et 222 al., 1995). This study demonstrated that FMD could be controlled and no further cases were 223 reported after a certain period of time. It would, therefore, be worthwhile to improve 224 management policies including destruction of infected beddings, regular use of disinfectants, 225 separation of uninfected animals from infected ones, and the quality of vaccines used. Protozoan 226 disease mortality of cattle from babesiosis was found to be higher in calves. This finding is 227 congruent with previous findings showing that babesiosis is prevalent in calves and only calves 228 (2.0%) died from babesiosis among the group of dairy cattle (Somavanshi, 1995). 229 230 The most important disease syndrome leading to deaths was pneumonia and a higher prevalence 231 of this disease was found in calves compared with adults. Similarly, other studies (Bhuller et al., 232 1985; Shimizu et al., 1987) also reported a higher proportion of calves dying from pneumonia. 233 Younger age, inadequate housing for newborn calves, climatic change, and lack of care might 234 contribute to the prevalence of pneumonia among calves. Enteritis is of particular importance as 235 a significant number of cattle died from this syndrome. Among them, calves were more common 236 than other categories of animals. Calves are more dependent on milk, milk replacer, and are 237 incapable of digesting all kinds of food materials. Overfeeding, sudden change in diet, poor 238 hygienic condition of feeding utensils, and calf barns might be the cause of death from enteritis. 239 Pneumonia and enteritis are multifactorial diseases that are closely associated with each other 240 (Debnath et al., 1995; Tikku, 1986). It has been suggested that common factors related to either 241 management techniques or specific agents predispose calves to both syndromes (Waltner-Toews 242 et al., 1986). Deaths caused by malnutrition were also more common in female calves. 243 Alternatively, a previous study (Mulei et al., 1995) reported that mortality caused by 13 14 244 malnutrition is more common in male calves. On the CCBDF it is not uncommon to find 245 emaciated and neglected calves. Particularly when the feed supply is not always adequate for all 246 animals or an incorrect proportion of feed ingredients is given to the animals. Debility was also 247 high in female calves. Lack of vegetation, unsuitable and insufficient food, and the prevalence of 248 other diseases might be the cause of debility. The majority of cattle that died from hairballs were 249 calves, but a few cases of cows and heifers were also found. Hairballs in calves might originate 250 from ingestion of hair during periods of excessive licking, and in adult animals is usually caused 251 by itchy skin conditions. Ingestion of hair is usually associated with persistent suckling of 252 penmates (Cockrill et al., 1978). Other studies (Abutarbush et al., 2006; Abutarbush et al., 2004) 253 also reported on the mortality of cattle from hairballs. 254 255 Conclusion: Among the risk factors associated with mortality, age of the animal was found to be 256 the most important followed by breed and season. Greater attention should be paid to the time of 257 colostrum feeding, proper timing and management of calves, and hygiene of calf barns. Because 258 of the complex nature of dairy management systems, a variety of causes are responsible for 259 diseases and deaths of dairy cattle. 260 261 References 262 Abutarbush, S.M. and Naylor J.M. (2006). Obstruction of small intestine by a trichobezoar in 263 cattle:15 cases (1992-2002). J. Am. Vet. Med. Assoc. 229: 1627-1630. 264 265 Abutarbush, S.M. and Radostits O.M. (2004). Obstruction of the small intestine caused by a 266 hairball in 2 young beef calves. Can. Vet. J. 45: 324-325. 14 15 267 268 Bhuller, M.S. and Tiwana M.S. (1985). Factors affecting mortality among buffalo calves. Indian 269 J. Anim. Sci. 55: 599-601. 270 271 Brenner, J., Orgad, V., Burd P. and Gat Z. (1989). The influence of climate on the mortality of 272 young calves in two farming systems in Israel. Israel J. Vet. Med. 45: 32-42. 273 274 Cockrill, J.M., Beasley J.N. and Selph R.A. (1978). Trichobezoars in four Angus cows. 275 Veterinary Medicine and Small Anim Clin. 73: 1441-1442. 276 277 Debnath, N.C., Taimura, M.J.F.A., Sahaa, A.K., Ersaduzaman, M., Helaluddin, M., Rahman, 278 M.L., Roy, D.K. and Islam M.A. (1995). A retrospective study of calf losses on the central 279 dairy cattle breeding station in Bangladesh. Prev. Vet. Med. 24: 43-53. 280 281 Debnath, N.C., Sil, B.K., Selim, S.A., Prodhan, M.A.M. and Howlader, M.M.R. (1990). A 282 retrospective study of calf mortality and morbidity on smallholder traditional farms in 283 Bangladesh. Prev. Vet. Med. 9: 1-7. 284 285 Islam, S.K.M.A., Hoque, M.A., Alam, M.R., Hassan, M.M. and Rahman, M.A. (2006). A Cross- 286 sectional study on production performance of stall fed dairy cattle at Central cattle Breeding 287 Station (CCBS), Savar, Dhaka, Bangladesh. Bangl. J. Vet. Med. 4: 61-63. 288 15 16 289 Islam, S.S., Ahmed, A.R., Ashraf, A., Khanom, N. and Ahmed, M.B. (2005). Causes and 290 Consequences of Calf Mortality in a Dairy Farm of Bangladesh. J. Anim. Vet. Adv. 4: 260-264. 291 292 Martin, S.W., Schwabe, C.W. and Franti, C.E. (1975). Dairy calf mortality rate: characteristics of 293 calf mortality rates in Tulare County, California. Am. J. Vet. Res. 36: 1099-1104. 294 295 Mitra, J., Das, D.K., Das, U. and Bardhan, G. (2005). Tubercle in lung of a crossbred cow. 296 Indian J. Anim. Health. 44: 77-78. 297 298 Mulei, C.M., Gitau, G.K. and Mbuthia, P.G. (1995). Causes of calf mortality in Kabete area of 299 Kenya. Onderstepoort J. Vet. Res. 62: 181-185. 300 301 Oluokun, S.B. and David-West, K.B. (1988). Analytical models of the national herd:Factors 302 controlling calf-mortality and their effects on rural economy. Acta Vet. Scand. 84: 419-422. 303 304 Prasad, S, Ramachandran, N. and Raju, S. (2004). Mortality patterns in dairy animals under 305 organized herd management conditions at Karnal, India. Trop. Anim. Health Prod. 36: 645-654. 306 307 Ramesh, P.S. (2007). Retrospective epidemiologic study of cattle diseases in central cattle 308 breeding and dairy farm, Savar, Dhaka. Bangladesh Agricultural University, 309 Mymensingh. MS thesis in Veterinary Medicine. 40–43. 310 16 17 311 Shimizu, T. and Nagatomo, H. (1987). Current status of calf diseases in japan. Bulletin of the 312 Faculty of Agriculture, Miyazaki University, 34: 320-336. 313 314 Somavanshi, R. (1995). Mortality pattern in a closed herd of dairy cattle in sub-temperate hilly 315 region. Indian Vet J. 72: 528-530. 316 317 Sultana, M., Ahad, A., Biswas, P.K., Rahman, M.A. and Barua, H. (2008). Black Quarter (BQ) 318 Disease in Cattle and Diagnosis of BQ Septicaemia Based on Gross Lesions and Microscopic 319 Examination. Bangladesh J Microbiol. 25: 13-16, 2008. 320 321 Thomsen, P.T., Kjeldsen, A.M., Sorensen, J.T. and Houe, H. (2004). Mortality (including 322 euthanasia) among Danish dairy cows (1990-2001). Prev. Vet. Med. 62: 19-33. 323 324 Thomsen, P.T., Kjeldsen, A.M., Sorensen, J.T., Houe, H. and Ersboll, A.K. (2006). Herd-level 325 risk factors for the mortality of cows in Danish dairy herds. Vet. Rec. 158: 622-626. 326 327 Tikku, A.K. (1986). Causes of calf mortality in Chanspa farm. Indian Vet. J. 63:690-692. 328 329 Waltner-Toews, D., Martin, S.W. and Meek, A.H. (1986). Dairy calf management, morbidity 330 and mortality in Ontario Holstein herds. Association of management with morbidity. Prev. Vet. 331 Med. 4: 137-158. 332 333 17