Lactoferrin, a glycoprotein with immunomodulatory and mast cell stabilising properties, in skin of horses suffering from Culicoides hypersensitivity

Lactoferrin (LF), a glycogen of the transferrin family with anti-bacterial and immunomodulatory properties, is expressed in various secretions and tissues. Cutaneous LF serves as a mast cell stabilising compound, modulates T cell activity and is found during IgE-mediated late phase reactions at allergen challenged sites. Culicoides hypersensitivity (CHS) in horses is a common IgE-mediated allergic dermatitis, characterised by an early and late phase cutaneous reaction upon allergen challenge. The aim of the study presented here was to examine whether LF mRNA expression in skin biopsies from horses affected by CHS prior to and 4h following intradermal challenge with a commercial C. nubeculosus extract is modified in comparison to skin biopsies from non-affected horses. In order to obtain reliable data, real time PCR was performed and genes of interest were normalized using three different housekeeping genes, beta-actin, GAPDH, beta-2-microglobulin. In comparison to non-affected horses, higher variation in LF mRNA levels both prior to and post-intradermal challenge with C. nubeculosus extract was seen in horses affected by CHS. However, the statistical analysis demonstrated that LF mRNA expression was not significantly different between CHS affected and non-affected horses prior to intradermal challenge with C. nubeculosus extract. Intradermal injection of C. nubeculosus extract did not result in local upregulation of LF mRNA at 4h post-injection. LF mRNA expression was therefore not significantly different pre- or post-intradermal challenge with C. nubeculosus extract in either group. Our data indicate that clinically normal skin of horses affected by CHS is not characterized by modified maintenance levels of LF mRNA. In contrast to human skin allergen challenged sites, LF mRNA levels in horses affected by CHS are not significantly different to that of control sites at 4h post-injection of C. nubeculosus extract.     

Are government veterinary paraprofessionals serving the poor? The perceptions of smallholder livestock farmers in Western Kenya

The liberalization of clinical veterinary services in Kenya introduced new service providers into the animal health service sector. This study examines the perceptions of livestock farmers regarding these service providers and analyses the factors that influence their choice of alternative service providers in Kakamega County. The empirical analysis shows that private animal health assistants were perceived to provide better services than alternative providers because they are more accessible and offer services on credit. Results from a multinomial logit model reveal that more educated, wealthier, and older farmers are more likely to use government services. The study concludes that it is imperative to better target the poor and to integrate private service providers into government animal health programs.

     

Autosomic 27 Trisomy in a Standardbred Colt

A 19-month-old Standardbred colt was donated to the University of Pennsylvania School of Veterinary Medicine with a suspicion of intersexuality. The anal−genital distance and penis were normal, and there was no evidence of intersexuality, but the colt was bilaterally cryptorchid. Several aspects of the colt's behavior appeared unusual, including general temperament and behavior described as sympathetically dull and affable. With herd mates, the colt appeared slow to perceive or to learn the usual intraspecies social cues. An atypical gait characterized by intermittent unnatural shuffle of the hind limbs, sliding them along in short rhythmic strides for 3 to 10 seconds at a time was noted at times when a horse might normally transition from a slow walk to a fast walk or a slow trot. Occasionally the colt exhibited slight protrusion of the tongue through the teeth and lips with jaw movements and smacking of the tongue against the teeth as if struggling to retract the tongue to the normal position. Evaluation of the karyotype combined with fluorescent in situ hybridization (FISH) revealed an abnormal male karyotype showing trisomy of chromosome 27 (65, XY + 27). The colt was euthanized at 24 months of age, and a necropsy revealed no significant abnormalities. This case of trisomy was not associated with developmental abnormalities described in other rare reports of trisomy in horses; however, some features were strikingly similar to that of humans with trisomy 21. FISH was demonstrated to be an excellent method for correct identification of equine chromosomes.     

How molecular methods change our views of FeLV infection and vaccination

FeLV was discovered 40 years ago and vaccines have been commercially available for almost two decades. So far, most FeLV pathogenesis and vaccine studies were conducted assaying parameters, such as virus isolation and antigen detection. Accordingly, regressive infection was characterized by transient or undetectable viremia, while persistent viremia is typically observed in cats with progressive infection. Using real-time polymerase chain reaction assays, the spectrum of host response categories to FeLV infection was recently refined by investigating proviral and plasma viral RNA loads. Cats believed to be immune to FeLV infection were found to turn provirus-positive after virus exposure. Moreover, efficacious FeLV vaccines were found unable to prevent provirus-integration and minimal viral replication. Remarkably, no difference was found in initial proviral and plasma viral RNA loads between cats with different infection outcomes. Only subsequently, the infection outcome is associated with FeLV loads. FeLV provirus was found to persist for years; reoccurrence of viremia and disease development was observed in some cats. Thus, aviremic provirus-positive cats are FeLV carriers and, following reactivation, may act as an infection source. However, integrated viral DNA may also be essential for solid protection and long-lasting maintenance of protective immunity. In conclusion, real-time TaqMan PCR and RT-PCR assays are highly sensitive and specific. They yield a more sensitive measure for FeLV exposure than antigen detection, virus isolation or immunofluoresence assays. We recommend the use of real-time PCR assays to identify FeLV exposed cats, particularly in catteries, and investigate obscure clinical cases that may be FeLV-associated. The use of sensitive molecular methods will contribute to a more in-depth understanding of the FeLV pathogenesis.     

Reference Values on Serum Biochemical Parameters of Brazilian Donkey (Equus asinus) Breed

Seventeen serum biochemical variables were determined in 40 donkeys of the Brazilian breed (34 females and 6 males) aged from 3 to 19 years. Mean ± standard deviation (SD) (minimum–maximum) values, obtained by automated analysis, were as follows: glucose, 58.35 ± 10.40 (44.00–90.00) mg/dL; cholesterol, 88.41 ± 9.86 (73.58–124.26) mg/dL; serum protein, 6.82 ± 0.40 (6.00–7.52) g/dL; albumin, 3.13 ± 0.21 (2.65–3.69) g/dL; creatinine, 1.80 ± 0.14 (1.51–2.19) mg/dL; urea, 24.25 ± 5.37 (14.12–34.39) mg/dL; lactate, 20.10 ± 4.58 (12.99–33.47) mg/dL; aspartate aminotransferase (AST), 295.81 ± 62.79 (173.71–466.07) IU/L; creatine kinase (CK), 158.00 ± 76.94 (51.69–440.33) IU/L; γ-glutamil-transferase (GGT), 45.82 ± 13.34 (26.17–86.38) IU/L; lactate dehydrogenase (LDH), 576.02 ± 156.32 (213.53–1162.81) IU/L; alkaline phosphatase (AP), 345.36 ± 65.90 (227.25–490.16) IU/L; calcium (Ca), 8.54 ± 0.18 (8.19–8.90) mg/dL; phosphorus (P), 2.76 ± 0.38 (1.99–3.97) mg/dL; chloride (Cl), 106.05 ± 3.20 (99.00–112.00) mEq/L; sodium (Na), 121.50 ± 4.14 (116.00–132.00) mEq/L; and potassium (K), 3.70 ± 0.42 (2.80–4.40) mEq/L. Comparisons of biochemical ranges obtained for the Brazilian donkey breed with reference ranges for other donkey breeds suggested that most values were similar. Biochemical values determined in the present study serve as reference ranges for donkey populations and can be used for health control and diagnosis of diseases.     

Frequencies of feline blood types in the Rio de Janeiro area of Brazil

Background: The distribution and frequency of blood types in cat populations vary according to geographic region and breed. Frequencies of feline blood types in Rio de Janeiro city, as well as in other Brazilian areas, are unknown, and the risk of unmatched transfusions and neonatal isoerythrolysis has not been estimated. Objectives: The purpose of this study was to determine the frequency of feline blood types in the area of Rio de Janeiro, Brazil. Methods: EDTA blood samples were obtained from 172 nonpedigreed domestic shorthair (DSH) cats (92 female, 80 male, 3 months-20 years old) in different sites of Rio de Janeiro city. Blood typing was performed by agglutination assays using Triticum vulgaris lectin and feline anti-A serum. The hemagglutination results for type B and AB cats were confirmed by high-performance thin-layer chromatography (HPTLC) of erythrocyte membrane gangliosides. Results: The majority (163/172, 94.8%) of cats were type A, 2.9% were type B, and 2.3% were type AB. High-titer anti-A serum agglutinated RBCs from all cats in type A and type AB blood groups, with 3+ to 4+ agglutination. The probability that a type A cat would receive type B or AB blood in a first random transfusion was calculated as 2.25% and 2.20%, respectively. HPTLC analysis of glycolipids yielded a chromatographic profile characteristic of feline gangliosides for all blood groups. Conclusions: These results indicate a high prevalence of type A cats in Rio de Janeiro, Brazil, and a low frequency of type B and AB cats, consistent with what has been observed for DSH cats in other regions of the world.     

Prevalence of Ehrlichia canis infection in thrombocytopenic dogs from Rio de Janeiro, Brazil

BACKGROUND: Infection with Ehrlichia canis causes a highly variable, multisystemic disease in dogs. Nevertheless, many clinicians in Rio de Janeiro, Brazil, use the presence of only thrombocytopenia to make a presumptive diagnosis of E canis infection. OBJECTIVE: The objective of this study was to determine the prevalence of E canis in thrombocytopenic dogs from Rio de Janeiro, Brazil, using polymerase chain reaction (PCR). METHODS: Following DNA extraction of whole blood samples from 226 dogs, PCR assays were done using primers for rickettsial DNA (including Ehrlichia spp, Anaplasma platys and A phagocytophilum) and using E canis-specific primers (16S rRNA gene). Dogs were grouped as thrombocytopenic and nonthrombocytopenic based on platelet counts. The null hypothesis that there was no difference in the prevalence of E canis in these groups was rejected at P<.05. RESULTS: Thirty-six (32.1%) of the thrombocytopenic dogs and 4 (3.5%) of the nonthrombocytopenic dogs were positive for rickettsial gene sequences (P<.0001). Further, 30 (26.8%) of thrombocytopenic dogs and 4 (3.5%) nonthrombocytopenic dogs were positive for E canis-specific gene sequences (P<.0001). CONCLUSIONS: Although the prevalence of E canis infection was higher in thrombocytopenic dogs, less than one third of these dogs had demonstrable E canis infection. Thus, thrombocytopenia is not specific for the detection of E canis infection and should not be used solely to establish a diagnosis of canine ehrlichiosis, even in a geographic area with relatively high disease prevalence.     

Seroprevalence of anaplasmosis among cattle in Switzerland in 1998 and 2003: no evidence of an emerging disease

Anaplasma marginale infection in Europe has been limited to the Mediterranean and eastern countries, to Austria and to very sporadic cases in Switzerland. There are no reports of its occurrence in the countries north of Switzerland. A severe outbreak of anaplasmosis in August 2002 in a cattle farm in the canton Grisons, Switzerland, north of the Alps, with more than 300 cattle that had to be culled, came unexpected and gave reason to hypothesize presence of an increased yet undetected prevalence of A. marginale in Switzerland. Randomly selected bovine serum samples collected in 1998 and 2003 were tested using a competitive inhibitory ELISA (cELISA) to test the hypothesis. Our validation of the diagnostic sensitivity and specificity of this test, done in the outbreak herd, yielded 99.2 and 83.3%, respectively, probably underestimating the true specificity. The true seroprevalence of anaplasmosis in Swiss cattle determined by cELISA was likely to be zero with upper 95% confidence limits of 2.49% in the canton Grisons and 1.17% in the rest of Switzerland, respectively, in 1998. For 2003, these estimates were even lower. There was no significant difference in apparent prevalences between 1998 and 2003. In search of a possible reservoir, three chamoises out of 46 free ranging wild ruminants from the Swiss National Park, Grisons, tested positive in the cELISA. This reaction is in accordance with A. marginale or a cross reacting agent such as Anaplasma ovis. From our results we conclude that the hypothesis of an increased prevalence of anaplasmosis in cattle in Switzerland must be rejected.     

Use of kit internal tandem duplications to establish mast cell tumor clonality in 2 dogs

Mast cell tumor (MCT) is one of the most common tumors of dogs. Some affected dogs develop multiple cutaneous tumors in various locations over months to years. In these cases, it is not clear whether the tumors have arisen de novo, or if each tumor represents a recurrence of the previously excised original tumor (ie, distant metastasis). We used the presence of an internal tandem duplication (ITD) in c-kit to demonstrate that in 2 dogs with recurrent cutaneous MCT that had developed over 1-2 years, each recurrent MCT tumor possessed an identical ITD when compared to the original MCT, indicating that the multiple tumors were clonal in origin. This study demonstrates that similar to the situation in humans, specific somatic mutations identified in oncogenes found in canine neoplasms can be used to provide evidence of tumor clonality.