What is new in the Wolbachia/Dirofilaria interaction?

Presence of transovarially-transmitted endosymbiontic Wolbachia bacteria in Dirofilaria immitis, and in other filariae of man and animals, presents a new paradigm for our understanding of pathogenesis, treatment and diagnosis of filarial infections. Many of the basic biological characteristics of Wolbachia have yet to be elucidated, but the results obtained to date suggest that canine or the feline hosts can be exposed to D. immitis Wolbachia when larvae, or adult worms, are killed; when Wolbachia are expulsed, with the deposition of microfilariae, from the uterus of the females; and possibly through the excretory system of both male and female worms. The two organs that have the greatest potential of being affected by the Wolbachial metabolic products/antigens released from the adult worms are the lungs and the kidneys. Population of Wolbachia in D. immitis is polymorphic. The life cycle of Wolbachia is complex and may consist of two reproductive modes: multiplication of the bacillary forms by binary fission and by a more complex mode which resembles the Chlamydia-like cycle that consists of three morphological stages: a small, dense body, an intermediate stage with a dense inclusion, and a bacillary form which represents the final product of development and maturation of the small, dense body. The Chlamydia-like cycle offers a potential survival strategy for the Wolbachia by producing more progeny than multiplication by binary fission, and appears to be more active during growth and development of embryos and of the larvae. The small, dense bodies may be the infectious forms responsible for the spread of Wolbachia through the canalicular system, within the lateral chords of filariae. An amorphous membrane that lines the perienteric surface of the body wall may represent a physical barrier that limits the spread and movement of Wolbachia to the perienteric surface of the lateral chords. Wolbachia in D. immitis may also offer therapeutic and diagnostic possibilities. Elimination of Wolbachia by chemotherapy, and the suppressive effect of aposymbiosis on embryonic development of D. immitis, may have potential application for control (sterilization of female worms) and treatment of dirofilariasis. However, the three stages in the life cycle of Wolbachia may be antigenically different and each stage may have a different susceptibility to therapeutic agents. Persistence of dormant small, dense bodies after treatment would allow the Wolbachia to re-establish once the conditions for development would become favorable. Detection of Wolbachial antigens provides an attractive diagnostic possibility to identify D. immitis early in the infection. Further studies on Wolbachia of filariae, including those of D. immitis, will undoubtedly reveal additional information that can be applied towards treatment, diagnosis, and control of filarial infections.     

Th1 response in BALB/c mice immunized with Dirofilaria immitis soluble antigens: a possible role for Wolbachia?

The immune response to filarial infection has been shown to be of both the Th1 and Th2 types. Studies aimed at developing immunization strategies against Dirofilaria immitis infection in dogs have shown that protection against larval challenge is of the Th2 type and that several proteins are recognized by immunized or infected animals. The bacterial endosymbiont Wolbachia, harbored by many filarial species including D. immitis, has recently been shown to interact with the host immune system. Specific antibodies to the Wolbachia recombinant surface protein (WSPr) have been observed in cats infected with D. immitis. In this work the authors have determined cytokine production and antibody response in BALB/c mice inoculated with soluble antigens from third stage larvae or from adult worms of D. immitis. Inoculated mice first produced IFN-gamma followed by a peak in IL-4. Specific antibodies to the Wolbachia protein WSPr were exclusively IgG2a, while antibodies against peptides derived from antigens of D. immitis were in the IgG1 and IgE subclasses. The cytokine response is thus similar to that reported for other filarial infection, where Th1 response shifts towards Th2. Antibody response indicates that Wolbachia may induce preferentially a Th1 response during filarial infection, while nematode antigens may be involved in Th2 response. There is thus an overall agreement with current opinions on the role of bacterial versus nematode molecules in driving the response towards the different directions.     

Epidemiological aspects of filariosis in dogs on the coast of Paraná state, Brazil: with emphasis on Dirofilaria immitis

The present study determined the prevalence and geographical distribution of Dirofilaria immitis and other filariae, from dogs in littoral areas of Paraná state, in Brazil. This survey spanned eight months, between 1998 and 1999, and was also designed to compare the efficacy of different tests for diagnosis of heartworm infection in that area. Blood samples were collected from 256 native-owned dogs distributed along the Paraná coastal area. Five diagnostic procedures were used: direct smear examination, the Knott's modified test, filtration assay, and two heartworm antigen detection kits. A follow-up imaging exam was performed to support the heartworm diagnosis. The imaging diagnosis included radiographic and ultrasonographic exams of six dogs that had positive results for the heartworm antigen detection kits, but showed different microfilarial burdens. The presence and severity of radiographic and ultrasonographic signs were compared with the results obtained in microfilariae detection and antigen tests. Diagnostic parasitology results indicated that 31.25% of the dogs were microfilaremic. Three different microfilariae were recovered: D. immitis, Dipetalonema reconditum, and the third (mf3) was not identified. D. reconditum was the species with the highest prevalence: 22.6%. In general, D. immitis prevalence was 5.47% (28.57% occult infections), but it varied along the coast and the range was from 0 to 20%. No correlation could be established between the overall scores for microfilarial counts (small or large numbers) and the severity of radiographic results or the likelihood of detecting filariae in the pulmonary artery using echocardiography. The finding of a different type of microfilaria (mf) suggested the existence of a third species in Paraná state, whose prevalence was 4.68%. These results show that to obtain a reliable diagnosis of heartworm infection, antigen detection kits are indicated. Knott's test or filtration should be performed to confirm microfilaremia and not for diagnosis of heartworm infection. Imaging tests support parasitology exams and add more about severity of infection. The northern areas, specially Guaraque?aba and Ilha das Pe?as, presented the highest number of heartworm-infected dogs.     

Pulmonary artery dissection causing haemothorax in a cat: potential role of Dirofilaria immitis infection and literature review

A 7-year-old male castrated domestic short-haired cat suddenly died. Gross examination revealed severe right-sided haemothorax with blood clots, four adult filarial nematodes in the blood clots and the caudal vena cava and haemorrhage dissecting into the tunica media of the right pulmonary artery. Histopathological investigation showed fibrosis of the tunica intima and disorganization/fragmentation of the elastic fibres accompanied by fibrous tissue deposition in the tunica media of both branches of pulmonary artery. Degenerative vasculopathy (intimal fibromuscular hyperplasia and medial hypertrophy/hyperplasia) involving pulmonary arteries was also observed. The polymerase chain reaction amplification and sequencing confirmed the identification of the parasite as Dirofilaria immitis. A diagnosis of pulmonary artery dissection with haemothorax and concomitant heartworm disease was formulated. Degenerative processes of the tunica media have been reported to cause pulmonary artery dissection in both humans and animals. Pulmonary artery remodelling induced by heartworms may be considered the underlying cause in the first case of feline pulmonary artery dissection, herein described.     

Seasonal or pathological findings? Morphofunctional characteristics of the equine endometrium during the autumn and spring transition

The deep anoestrous phase in winter is part of the anovulatory season in mares and is bordered by the autumn and spring transitional periods (ATP/STP). To define an annual time span for effective prognostic biopsy sampling, the aim of this study was to provide a morphofunctional characterization of the endometrium during ATP and STP. To outline both transitional periods, endometrial specimens were taken in September, October and November (n = 76) as well as February, March and April (n = 184) with the requirement of a detailed clinical documentation. Tissue samples were examined histologically with special emphasis on the functional endometrial morphology. Additionally, an immunohistochemical evaluation was performed on selected specimens regarding the expression of oestrogen receptor α, progesterone receptor and Ki67‐antigen. An absent to low endometrial activity was ascertained in more than 60% of all specimens from late October onwards, whereas a comparably lacking or low activity in STP was observed until early April. Approximately 30% (ATP) to 22% (STP) of all samples exhibited a predominantly “irregular” endometrial differentiation. During the transitional periods, the clinically evaluated ovarian status (transrectal palpation, transrectal ultrasonography and/or serum progesterone and oestrogen analyses) and the endometrial functional morphology were in accordance with approximately 70% of all cases. The expression of steroid hormone receptors and Ki67‐antigen was generally low. Given that endometrial maldifferentiations were frequently found during ATP and STP, its occurence might display a characteristic and physiological feature of the transitional periods. Regarding the functional endometrial morphology, a diagnostic biopsy sampling should therefore be performed between late April and before September.     

Is breed-specific legislation justified? Study of the results of the temperament test of Lower Saxony

Beginning in July 2000 legislation in Lower Saxony restricted the keeping of bull terriers, American Staffordshire terriers, pit bull terriers, and 11 other breeds. Exemption was possible after attending an especially developed test. The tests analyzed for breed predisposition for excessively aggressive signaling or aggressive behavior in inappropriate situations, differences in behavior between breeds, and factors differentiating biting from nonbiting dogs.The test results of 415 dogs were analyzed. The test consisted of a veterinary examination; a learning test; situations of dog-human, dog-environment, and dog-dog contact; and obedience. Escalation in aggressive behavior was scored using a scale of 1-7. No aggressive behavior (1 on the scale) was shown by 38.07% of the dogs, 61.69% showed aggressive behavior scored as 2 to 5 on the scale, and 0.24% bit without previous threatening signals (6 on the scale).Concerning a score of 1 on the scale, pairwise comparison (chi-square test) showed significant differences between bull terriers and American Staffordshire terriers (P = 0.004), pit bull terriers (P = 0.01), Doberman pinschers (P = 0.003), and rottweilers (P = 0.009). Concerning scores of 2 to 7 on the scale,, no significant differences were found.Ninety-five percent of the animals reacted appropriately in the test situation. Five percent displayed excessive aggressive signaling or aggressive behavior in inappropriate situations. These displays were associated with unusual movements and the dogs' apparent apprehension. Correlation between test results and owners jerking on the leash or misinterpreting their dogs' behavior and dogs trying to elude physical manipulation was found. No significant difference in behavior between breeds was detected.The results show no indication of dangerousness in specific breeds. Justification for specific breed lists in the legislation was not shown.     

Practice management: Is necessity the mother of contention?

Extract

During much of this century veterinary undergraduates were led to believe that because the practice of veterinary science is a profession, after graduation they should never treat clinical practice as a business. This notion was reinforced by their peers, especially. those outside clinical practice, and still exists today. With the dependence of rural practice on the supply and sale of remedies for viability, few practices have ever been run on purely professional lines in New Zealand; but until very recently a minimal level of business ability was adequate to maintain practice viability.     

Is the Function of the Porcine Sperm Reservoir Restricted to the Ovulatory Period?

The uterotubal junction (UTJ) and caudal isthmus are recognized as a functional pre-ovulatory sperm reservoir (SR). Spermatozoa are released from the SR in a complex and concerted action. However, whether this functionality is restricted only to the ovulatory period is still open to debate. Our study was aimed to analyze the presence of spermatozoa within the UTJ (SR), isthmus (ISTH) and ampulla (AMP) after laparoscopic intrauterine insemination (LIUI) either in the peri- (PERI) or post-ovulatory (POST) period or at mid cycle (MID). Each uterine horn of estrus synchronized gilts (n=12) was inseminated with 20 ml sperm (29.5×10⁶ cells/ml). Oviducts were recovered 7 h after LIUI and separated into the UTJ, ISTH and AMP, and sections were flushed with 10 ml PBS+EDTA solution. After centrifugation, the sperm pellet was evaluated by Čeřovský staining. The median sperm numbers in the PERI, POST and MID groups were 578, 171 and 789 in the UTJ; 545, 233 and 713 in the ISTH; and 496, 280 and 926 in the AMP, respectively, and there were differences between the POST and MID groups (P<0.05) but not between the oviductal sections of each group (P>0.05). Compared with the MID group, the percent of intact sperm cells was higher (P<0.01) in the PERI and POST groups (32.8 vs. 66.4 and 76.8%). Also, the percentages of aberrations in the acrosome and tail were higher (P<0.05) in the MID group. Based on this, it can be assumed that the sperm reservoir is active during different phases of the estrus cycle. However, the mid-cycle oviduct environment considerably impairs sperm cell quality.     

Is the use of formulae a reliable way to predict the accuracy of genomic selection?

We studied four formulae used to predict the accuracy of genomic selection prior to genotyping. The objectives of our study were to investigate the impact of the parameters of each formula on the values of accuracy calculated using these formulae, and to check whether the accuracies reported in the literature are in agreement with the formulae. First, we computed the marginal distribution of accuracy (by integration) for each parameter of all four formulae: heritability h², reference population size T, number of markers M and number of effective segments in the genome M_e. Then, we collected 145 accuracies and corresponding parameters reported in 13 publications on genomic selection (mainly in dairy cattle), and performed analysis of variance to test the differences between observed and predicted accuracy with effects of formulae and parameters. The variation of accuracy for different values of each parameter indicated that two parameters, T and M_e, had a significant impact and that considerable differences existed between the formulae (mean accuracies differed by up to 0.20 point). The results of our meta‐analysis showed a big formula effect on the accuracies predicted using each formula, and also a significant effect of the value obtained for M_e calculated from N_e (effective population size). Each formula can therefore be demonstrated to be optimal depending on the assumption used for M_e. In conclusion, no rules can be applied to predict the reliability of genomic selection using these formulae.