Detection of Avian Influenza Virus H5N1 in Horses at Assiut Governorate, Egypt

The highly pathogenic H5N1 is a major avian pathogen that intensively affects the poultry industry in Egypt, even in spite of the adoption of vaccination strategy. The virus is currently panzootic in Egyptian poultry populations and crosses species barriers to humans and animals. In February 2014, 15 horses at El-Fath center, Assiut, Egypt, started to show mild fever, dullness, restlessness, slight nasal discharge and cough. Two weeks later one of these horses died and another one became recumbent. This was associated with the spread of avian influenza cases in the backyard birds in the same area. Serum samples were collected from the diseased horses and from birds in the same area and examined by haemagglutination inhibition (HI) assay for detection of viral antibodies. At the same time, nasal swabs from horses and tracheal swabs from birds collected and examined by rapid antigen detection and quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) for detection of the virus. Serum samples of horses showed positive titer with HI in 13 out of the 14 diseased horses and the highest titer was 6-log2 in the recumbent one. Meanwhile, the result of HI for birds serum samples (450) were negative for 425 samples, which indicate absence of previous vaccination or infection, while the remaining 25 samples were from balady chickens showing HI titer 6-log2±0. 84 and those birds were previously vaccinated 4 weeks before collecting the serum. Avian influenza H5N1 virus could not be detected by using rapid antigen detection strips in the nasal swabs taken from the diseased horses, but could be detected in birds in 102/150 with antigen capture immune-assay (AC-EIA) common antigen type A, 95/150 with (AC-EIA) H5, 0/150 with (AC-EIA) H7 and 0/150 with (AC-EIA) H9 of examined samples. By qRT-PCR, H5N1 virus could be detected only in the nasal swab of the recumbent horse, and in 138/150 tracheal bird swab. In conclusion, Assiut city in Egypt is a disease endemic area where the probability of intimate contact between infected backyard birds and horses is high. Therefore, the disease may be transmitted to these horses from aerosol exposure of infected birds' droppings or contaminated feeds and water or because of direct contact with infected birds. However, the moderate severity of the H5N1 in equine may be responsible for the recovery of most of the diseased horses without further complications

Reference Values for Serum Biochemical and Hematological Constituents in Lactating Pregnant Buffaloes

Serum biochemical and hematological reference values are used to establish normality and to diagnose disease and physiological alterations. Up till now there are no reference values for different blood biochemical and hematological variables in lactating pregnant buffaloes, consequently the current study considered the first one that established those values in lactating pregnant water buffaloes (Bubalus bubalis). Animals were examined at buffalo farms that belong to Assiut Governorate, Egypt. A total of 148 pregnant lactating buffaloes (5-10 years old) were examined, of these, 20 buffaloes did not meet the selection criteria and excluded from study. The remained animals (N.=128) were clinically healthy and included in the study. A total of 55 hematological and serum biochemical variables were measured in blood of lactating buffaloes. Mean reference values, reference intervals (2.5 and 97.5 percentiles) and their 90% confidence intervals, for the measured hematological and serum biochemical constituents were statistically calculated. Data for the serum biochemical and hematological variables were presented in tables and compared with previously published data. The established reference values will be a useful guide for interpreting serum biochemical and hematologic data in lactating pregnant water buffaloes

Comparative Study on Reference Values for Blood Constituents during Pregnancy in Buffaloes (Bubalus bubalis)

Reference values for buffaloes especially those at pregnancy are not yet established. The aim of this study was to establish serum biochemical and hematological reference values for water buffaloes (Bubalus bubalis) during pregnancy. In total 409 pregnant buffaloes were examined at buffaloes' farms that belong to Assiut Governorate at the mid of Egypt. Out of them, 107 buffaloes did not meet the selection criteria and were excluded from the study. The remained 302 clinically healthy buffaloes were classified according to the stage of pregnancy into two groups: Group I; included buffaloes till 6 months of pregnancy (No.=146). Group II; included buffaloes after 6 months of pregnancy (No.=156). Three types of samples were collected; serum samples for biochemical analysis, whole blood samples for hematological analysis and fecal samples for parasitological examination. A total of 55 blood variables were measured during this study. The 95% reference intervals for each serum biochemical and hematological constituents were calculated by removing the upper and lower 2.5% of the interval to give the 2.5 and 97.5 percentiles. The present study established the reference intervals for the investigated biochemical and hematological parameters in blood of pregnant buffaloes. Results revealed that most of the measured blood constituents were differed significantly during the period before and after 6 months of pregnancy in buffaloes. In conclusion, the established reference values will be a useful guide for interpreting serum biochemical and hematologic data in pregnant buffaloes

Reference Intervals for Rectal Temperature in Water Buffalo (Bubalus bubalis) Heifers

To the best of the authors knowledge, reference intervals for rectal temperature in water buffalo (Bubalus bubalis) has not been previously published. This study was undertaken to establish this reference interval for water buffalo heifers. Based on the statement of the International Federation of Clinical Chemistry, at least 120 values are necessary to obtain reliable estimates for reference intervals. A total number of 127 healthy buffalo heifers (1–2 years old) were selected based on a set of inclusion criteria. The health status of heifers was confirmed based on clinical examinations and laboratory analyses. Animals were examined at buffalo farms that belong to Assiut Governorate, Egypt. Rectal temperature was measured using a clinical thermometer. Three types of samples were collected: serum samples for biochemical analysis, whole blood samples for haematological analysis and faecal samples for parasitological examination. The 95% reference intervals were calculated by removing the upper and lower 2.5% of the interval to give the 2.5 and 97.5 percentiles. Confidence intervals were calculated for the reference limit. The current study was able to establish reference intervals for rectal temperature in water buffalo heifers

Serum biochemical and haematological reference intervals for water buffalo (<i>Bubalus bubalis</i>) heifers

Based on a review of the literature, reference intervals for water buffalo (Bubalus bubalis) serum biochemistry and haematology have not previously been published. The current study was done to establish reference intervals for water buffalo heifers. The International Federation of Clinical Chemistry stated that at least 120 values are necessary to obtain reliable estimates for reference intervals. A total number of 127 clinically healthy buffalo heifers (1–2 years old) were included in the study. Animals were examined at buffalo farms that belong to Assiut Governorate, Egypt. Three types of samples were collected: serum samples for biochemical analysis, whole blood samples for haematological analysis and faecal samples for parasitological examination. Animals that fitted the inclusion criteria were included in the study. Biochemical analysis included serum total proteins, albumin, total globulins, alpha, beta and gamma globulin levels, and aspartate aminotransferase, alanine aminotransferase, gamma glutamyl transferase, creatine phosphokinase and lactate dehydrogenase activity. In addition to the above, serum creatinine, urea, total bilirubin, direct bilirubin, indirect bilirubin, sodium, potassium, chloride, magnesium, calcium, phosphorus, copper, zinc, iron, triglycerides, high density lipoprotein, low density lipoprotein, very low density lipoprotein, glucose levels and 20 haematological variables were measured. The 95.0% reference intervals were calculated by removing the upper and lower 2.5% of the interval for each serum biochemical constituent to give the 2.5 and 97.5 percentiles. Confidence intervals were calculated for each reference limit. Reference intervals from the current study were compared with established values for cows. The current study is as far as could be determined the first that establishes reference intervals for the serum biochemical and haematological parameters in water buffalo heifers