Quantitation of reticulated platelets in healthy dogs and in nonthrombocytopenic dogs with clinical disease

Background — Thrombocytopenia is a common disorder in dogs and development of an objective diagnostic assay to measure platelets newly released from bone marrow into the blood would provide a noninvasive way to predict megakaryocytopoiesis. Reticulated platelets are newly released platelets with increased concentrations of RNA that can be detected by flow cytometric analysis of blood stained with thiazole orange (TO).
Objectives — The goals of this study were to establish a reproducible method to quantitate reticulated platelets in dogs, to establish a reference interval for reticulated platelet percentages in healthy dogs, and to determine whether the percentage of reticulated platelets was nonspecifically increased in nonthrombocytopenic dogs with clinical disease.
Methods — Blood samples were obtained from healthy dogs and from nonthrombocytopenic dogs presented for a variety of disorders. An aliquot of whole blood was stained with TO and a phycoerythrin-labeled monoclonal antibody to platelet CD61, then analyzed by flow cytometry.
Results — The coefficients of variation were 7.8% to 15.6% (intra-assay precision) and 6.1% to 19.5% (interassay precision). Overnight storage for 18 to 26 hours, under variable conditions, resulted in an increase in the percentage of platelets staining with TO. The reference interval for reticulated platelets in the healthy control group was 0–4.3% (0–12,095/μL). No significant differences were found in the mean percentage of reticulated platelets or absolute concentration of reticulated platelets between control and affected dogs.
Conclusions — These studies demonstrate a reliable, noninvasive diagnostic assay for measurement of reticulated platelets in whole blood and provide a baseline for assessment of the clinical utility of the assay.     

Myopathy and alterations in serum 3-methylhistidine in dogs with liver disease

Liver disease can influence the metabolism of various other organs. Regarding the influence of liver diseases on muscles, only a few studies done on people exist. The goal of our study was to investigate the influence of liver diseases on muscles in dogs. Twenty-eight dogs with different liver diseases were investigated in this study. The diagnosis of muscle alteration was based on electromyography (EMG), creatine kinase serum activity, 3-methylhistidine serum concentration and a muscle biopsy in some cases. Our results suggest that liver diseases in dogs can be accompanied with muscle alteration. 3-Methylhistidine serum concentration as a new parameter for muscle destruction in dogs was significantly increased compared to clinical healthy dogs and was comparable to those concentrations in dogs with histologically confirmed myopathy of different types. The differentiation of the liver diseases into severe hepatitis, moderate hepatitis and liver tumours showed a significant elevation of 3-methylhistidine serum concentration in cases of liver tumours (P=0.03) and a tendency in cases of severe hepatitis (P=0.07). Based on our study we can conclude that liver diseases have an influence on muscles in dogs and 3-methylhistidine could be a useful parameter for muscle destruction.     

SONOGRAPHIC EVALUATION OF EXPERIMENTAL BILE DUCT LIGATION IN THE DOG

The common bile duct was surgically ligated in five normal adult dogs. Ultrasonographic examinations of the gallbladder and biliary system were performed after duct ligation at intervals of 24 hours. The sequence of biliary system dilation was from the common duct to the peripheral intrahepatic ducts. Common duct enlargement was evident in 24–48 hours, while peripheral biliary duct dilation was recognized by five to seven days after obstruction. When compared with hepatic and portal veins, dilated biliary ducts were more tortuous and had irregular branching patterns. Gross pathologic changes were correlated with ultrasonographic findings at seven, 14, 15, 18 and 21 days after obstruction.     

Restricted energy intake and elevated calcium and phosphorus intake for gilts during growth. IV. Characterization of metacarpal, metatarsal, femur, humerus and turbinate bones of sows during three parities

The effect of early nutrition on subsequent bone development was studied using gilts that had previously been fed ad libitum or 75% ad libitum intake and 100 or 150% National Research Council-recommended daily Ca and P from weaning to 100 kg. During the three-parity reproductive study, sows were fed a 14% protein diet. Metacarpals and metatarsals were taken from sows culled due to lameness or failure to breed and from sows after completing three parities. Femur and humerus articular cartilage and turbinates were described at necropsy. Metacarpals and metatarsals were heavier and tended to have thicker walls when sows were previously fed ad libitum or fed 150% Ca and P. Energy intake produced the greater response. Metacarpal breaking strength was greatest for sows previously fed ad libitum. Metatarsals were not affected by energy intake. Stiffness, Young's modulus of elasticity (YME) and flexural modulus for metacarpals and metatarsals were not affected by energy intake. Previously fed Ca and P intakes did not affect any of the mechanical bone characteristics. Metacarpals were heavier, had a greater breaking strength, were more elastic and exhibited slightly less resistance to bending than the metatarsals. The ether extract, ash, Ca and P content and the Ca:P ratio of metacarpals and metatarsals were not affected by previously fed energy or Ca and P intakes. The ether extract content tended to decrease and the ash, Ca and P content tended to increase with age. Articular cartilage and turbinate scores were not influenced by previously fed energy or Ca and P intakes. The YME and flexural modulus were the only bone characteristics that were even poorly correlated (average r = .25) with soundness scores, suggesting that high values result in poorer soundness scores. Energy and Ca and P intakes during growth and development had only minimal effects on bone characteristics and no apparent effect on structural soundness and longevity of sows kept for three parities.     

Putative human and avian risk factors for avian influenza virus infections in backyard poultry in Egypt

Highly pathogenic influenza A virus subtype H5N1 causes significant poultry mortality in the six countries where it is endemic and can also infect humans. Egypt has reported the third highest number of poultry outbreaks (n = 1084) globally. The objective of this cross-sectional study was to identify putative risk factors for H5N1 infections in backyard poultry in 16 villages in Damietta, El Gharbia, Fayoum, and Menofia governorates from 2010–2012. Cloacal and tracheal swabs and serum samples from domestic (n = 1242) and wild birds (n = 807) were tested for H5N1 via RT-PCR and hemagglutination inhibition, respectively. We measured poultry rearing practices with questionnaires (n = 306 households) and contact rates among domestic and wild bird species with scan sampling. Domestic birds (chickens, ducks, and geese, n = 51) in three governorates tested positive for H5N1 by PCR or serology. A regression model identified a significant correlation between H5N1 in poultry and the practice of disposing of dead poultry and poultry feces in the garbage (F = 15.7, p < 0.0001). In addition, contact between domestic and wild birds was more frequent in villages where we detected H5N1 in backyard flocks (F = 29.5, p < 0.0001).