Veterinary and public health aspects of Toxocara spp

Pet dogs and cats can play an important role in the transmission of zoonotic nematodes such as Toxocara canis and Toxocara cati, by excreting eggs directly into the human environment, without the involvement of vectors or intermediate hosts. Human toxocarosis remains a hazard despite the availability of highly effective anthelmintics for dogs and cats. A good understanding of the biology and epidemiology of these parasites, and the risk factors that lead to their transmission to humans is required for effective prevention strategies. In this respect, the maintenance of high quality continuing education for veterinarians and the provision of suitably presented information to pet owners are of priority importance. A closer collaboration between veterinary and public health professionals within the ‘One Health’ concept is also required.     

Prevalence of intestinal parasites and genotyping of Giardia intestinalis in pet shop puppies in east Japan

The current study examined the prevalence of intestinal parasites and genotypes of Giardia intestinalis in puppies from nine pet shops in east Japan. Fresh fecal samples from 1794 puppies (≦3 months old) were collected on one occasion. Giardia spp. was examined for specific coproantigen using ELISA kit (SNAP^®Giardia, IDEXX Laboratories, Inc., USA). Other intestinal parasites were detected microscopically using the formalin-ethyl acetate sedimentation technique. Genotyping was determined for the random 29 stool samples identified as Giardia spp. positive using PCR and direct sequencing of the glutamate dehydrogenase (gdh) gene. Overall prevalence of protozoan Giardia spp. and Cystoisospora spp. revealed 23.4% and 11.3%, respectively. Prevalence of ascarids, Strongyloides spp. and hookworms were recorded 1.8%, 1.1% and 0.1%, respectively. Protozoan Giardia spp. and Cystoisospora spp., thus, represent important pathogens among pet shop puppies. All genotyped G. intestinalis isolates were belonged to assemblage C or D, identified as dog-specific genotypes. Zoonotic assemblage A and B were not demonstrated. The result suggests that the risk of zoonotic transmission of G. intestinalis from pet shops puppies to humans may be quite low in Japan.     

Detection and identification of Toxocara canis DNA in bronchoalveolar lavage of infected mice using a novel real-time PCR

Toxocarosis is a zoonosis with worldwide distribution caused by Toxocara spp. of dogs and cats. In humans, diagnosis relies mainly on detection of parasite-specific antibodies. Although serological assays in current use have defined sensitivity and specificity, the problem of cross-reactivity still remains, particularly in areas of endemic polyparasitism. Microscopic detection of the parasite in tissue biopsies is not recommended for diagnosis because larvae can be difficult to locate, and finding the parasite eggs in faeces is not applicable since the larvae do not develop to the adult stage in the human host. In this study we describe a novel real-time PCR (‘Nemo-PCR’) that, in combination with DNA sequencing, allows the detection and identification of Toxocara canis and other nematodes in the Superfamily Ascaridoidea. Results indicate that this approach can detect Toxocara spp. DNA in bronchoalveolar lavage (BAL) of experimentally-infected mice. For diagnostic purposes further studies are necessary to evaluate this assay including testing human BAL fluid. The availability of such a direct assay would improve diagnosis of toxocarosis particularly for patients with pulmonary signs and symptoms.     

Bartonella spp. in cats from Buenos Aires,Argentina

In Argentina, data on the presence of members of the genus Bartonella is scarce. To increase knowledge about these zoonotic pathogens in this country, the presence and variability of Bartonella spp. was investigated in cats and dogs from Buenos Aires. Bartonella spp. was detected in 17.8% of cats, while all dogs tested negative by PCR and Reverse Line Blot. B. henselae was the most frequent species, being detected in 11.9% (14/101), while B. clarridgeiae was found in only 5.9% (6/101) of the cats. Afterwards, B. henselae isolates and positive blood samples were characterized by Multiple Locus Sequence Typing (MLST) and Multiple Locus Variable Number Tandem Repeats Analysis (MLVA). As result, four different MLST sequence types (ST) and eight MLVA profiles were identified. ST 1 was the most frequent variant found in cats, followed by ST 8. Interestingly, some of the MLVA profiles that were detected in this study have been previously associated with human disease, and represents a potential risk of infection. Veterinarians and physicians should consider the presence of these emerging pathogens in their diagnostic routine.     

Lungworm infections (Angiostrongylus vasorum, Crenosoma vulpis, Aelurostrongylus abstrusus) in dogs and cats in Germany and Denmark in 2003–2007

Faecal samples of 4151 dogs from Denmark, 958 dogs from Germany and 231 cats from Germany with clinical signs were examined for lungworm larvae using the Baermann funnel technique between 2003 and 2007. In total, 3.6% of Danish and German dogs shed lungworm larvae. In Denmark, patent infections of dogs with Angiostrongylus vasorum were more prevalent (2.2%) than those with Crenosoma vulpis (1.4%). In Denmark, the majority of A. vasorum- (98%) and C. vulpis-infected (80%) dogs originated from Northern Zealand. The frequency of A. vasorum and C. vulpis infections in Danish dogs obviously decreased from 2003 to 2006. In Germany, canine faecal samples were found more frequently positive for C. vulpis than for A. vasorum larvae (2.4% and 1.2%, respectively). Lungworm-infected dogs originated mainly from southern and western Germany. Larvae of Aelurostrongylus abstrusus were detected in 5.6% of cats from Germany. Overall, a distinct seasonal pattern in the detection of infected dogs was apparent for A. vasorum in Denmark and C. vulpis in Germany. The relatively high number of lungworm-infected dogs and cats indicate that these parasitic diseases should be considered in differential diagnosis of cases of treatment-resistant respiratory/cardiopulmonary distress.     

Babesiosis in dogs and cats--expanding parasitological and clinical spectra

Canine babesiosis caused by different Babesia species is a protozoal tick-borne disease with worldwide distribution and global significance. Historically, Babesia infection in dogs was identified based on the morphologic appearance of the parasite in the erythrocyte. All large forms of Babesia were designated Babesia canis, whereas all small forms of Babesia were considered to be Babesia gibsoni. However, the development of molecular methods has demonstrated that other Babesia species such as Babesia conradae, Babesia microti like piroplasm, Theileria spp. and a yet unnamed large form Babesia spp. infect dogs and cause distinct diseases. Babesia rossi, B. canis and Babesia vogeli previously considered as subspecies are identical morphologically but differ in the severity of clinical manifestations which they induce, their tick vectors, genetic characteristics, and geographic distributions, and are therefore currently considered separate species. The geographic distribution of the causative agent and thus the occurrence of babesiosis are largely dependent on the habitat of relevant tick vector species, with the exception of B. gibsoni where evidence for dog to dog transmission indicates that infection can be transmitted among fighting dog breeds independently of the limitations of vector tick infestation. Knowledge of the prevalence and clinicopathological aspects of Babesia species infecting dogs around the world is of epidemiologic and medical interest. Babesiosis in domestic cats is less common and has mostly been reported from South Africa where infection is mainly due to Babesia felis, a small Babesia that causes anemia and icterus. In addition, Babesia cati was reported from India and sporadic cases of B. canis infection in domestic cats have been reported in Europe, B. canis presentii in Israel and B. vogeli in Thailand. Babesiosis caused by large Babesia spp. is commonly treated with imidocarb dipropionate with good clinical response while small Babesia spp. are more resistant to anti-babesial therapy. Clinical and parasitological cure are often not achieved in the treatment of small Babesia species infections and clinical relapses are frequent. The spectrum of Babesia pathogens that infect dogs and cats is gradually being elucidated with the aid of molecular techniques and meticulous clinical investigation. Accurate detection and species recognition are important for the selection of the correct therapy and prediction of the course of disease.     

Virulence factors and genetic variability of uropathogenic Escherichia coli isolated from dogs and cats in Italy

In this study, the association between virulence genotypes and phylogenetic groups among Escherichia (E.) coli isolates obtained from pet dogs and cats with cystitis was detected, and fingerprinting methods were used to explore the relationship among strains. Forty uropathogenic E. coli (UPEC) isolated from dogs (n = 30) and cats (n = 10) in Italy were analysed by polymerase chain reaction (PCR) for the presence of virulence factors and their classification into phylogenetic groups. The same strains were characterized by repetitive extragenic palindromic (REP)- and enterobacterial repetitive intergenic consensus (ERIC)-PCR techniques. We found a high number of virulence factors such as fimbriae A, S fimbriae (sfa) and cytotoxic necrotizing factor 1 (cnf1) significantly associated with phylogenetic group B2. We demonstrated a high correlation between α-hemolysin A and pyelonephritis C, sfa, and cnf1 operons, confirming the presence of pathogenicity islands in these strains. In addition, UPEC belonging to group B2 harboured a greater number of virulence factors than strains from phylogenetic groups A, B1, and D. REP- and ERIC-PCR grouped the UPEC isolates into two major clusters, the former grouping E. coli strains belonging to phylogenetic group B2 and D, the latter grouping those belonging to groups A and B1. Given the significant genetic variability among the UPEC strains found in our study, it can be hypothesized that no specific genotype is responsible for cystitis in cats or dogs.     

Genotype identification and prevalence of Giardia duodenalis in pet dogs of Guangzhou, Southern China

Giardia duodenalis is a flagellated parasite and is considered one of the most common causes of protozoal diarrhea in both humans and animals worldwide. This paper represents the first study of the prevalence of G. duodenalis in pet dogs in Guangzhou, China. Faecal samples (209 specimens) were obtained from young (<6 months old), adult (6 months to 3 years) and elder dogs (>3 years old). 8.61% (18/209) faecal samples were recorded positive using microscopy examination, and 11.00% (23/209) using PCR. The prevalence was significantly higher in diarrheic dogs (26.31%) compared with non-diarrheic dogs (5.10%), while it was higher in young (25.58%) than both adult (7.37%) and elder (7.04%) dogs and the difference was statistically significant (P<0.05). The prevalence in male dogs 11.30% (13/115) was higher than females 10.87% (10/92), and in suburban dogs (12.15%) higher than urban 9.80%, but the difference was not statistically significant (P>0.05). Sequence analysis of the 23 PCR-positive samples revealed the presence of Assemblage D (18/23), and zoonotic Assemblage A (5/23). The present investigation reported a high infection rate of G. duodenalis in pet dogs, especially in young dogs. Genotypic characterization demonstrated that the zoonotic Assemblage A was found, a fact that poses a potential risk of G. duodenalis transmission from pet dogs to humans. It is suggested that pet owners should take appropriate hygiene measures to prevent and control giardiasis in this region.     

Toxocara spp. infections in paratenic hosts

The zoonotic roundworms Toxocara canis and T. cati are not only present worldwide in their definitive hosts; they also frequently occur in other animal species, including humans. In those so-called paratenic hosts, the larvae do not develop into the adult stage, but rather migrate throughout the somatic tissue and persist as infectious L3 stage for extensive periods. Those arrested larvae may lead to severe inflammatory reactions and consequently to a wide range of pathological and clinical manifestations. However, the infected paratenic hosts also constitute a potential source of infection for the definitive hosts or humans who may also function as paratenic hosts. In the present review, current knowledge of larval migration in a variety of possible paratenic hosts is summarized including variations of migration routes and susceptibilities. Furthermore, information about the clinical and pathological changes for the presented species and possible consequences of the somatic migration of larvae, i.e. the resulting tissue damage as well as adverse host reactions to arrested larvae are reviewed. There are still many questions unanswered regarding larval behaviour in hosts other than their definitive host. Therefore, it is of great importance to continue further elaboration on the biology of Toxocara spp. to prevent further spreading of larvae in both the paratenic and the definitive host.     

Enteric parasites of free-roaming,owned, and rural cats in prairie regions of Canada

The objective of this study was to determine prevalence, intensity, and zoonotic potential of gastrointestinal parasites in free-roaming and pet cats in urban areas of Saskatchewan (SK) and a rural region in southwestern Alberta (AB). Fecal samples were analyzed using a modified double centrifugation sucrose flotation to detect helminth eggs and coccidian oocysts, and an immunofluorescence assay to detect Giardia and Cryptosporidium. Endoparasite prevalence was higher in samples from rural AB cats (41% of 27) and free-roaming SK cats (32% of 161) than client-owned SK cats (6% of 31). Parasites identified using morphological and molecular techniques included Toxocara cati, Toxascaris leonina, Baylisascaris-type eggs, Eucoleus aerophilus, Taenia taeniaeformis, Isospora spp., Cryptosporidium spp., and zoonotic genotype A of Giardia duodenalis. This study demonstrates significant differences in endoparasite prevalence in feline populations, and the value of molecular techniques in fecal-based surveys to identify and determine parasite zoonotic potential.     

Prevalence and characterization of Staphylococcus aureus and Staphylococcus pseudintermedius isolated from companion animals and environment in the veterinary teaching hospital in Zambia,Africa

The Republic of Zambia consists of only one veterinary teaching school at the University of Zambia (UNZA) where students and veterinarians are exposed to many bacterial pathogens including Staphylococcus aureus (SA) and Staphylococcus pseudintermedius (SP). The aim of this study was the characterization and antimicrobial susceptibility profile of eleven SA and 48 SP isolates from the veterinary hospitals’ in- and outpatients and the environment. No isolate was resistant to cefoxitin by disk diffusion test and the corresponding resistance gene mecA was not found. In contrast, the resistance rates of SA to penicillin (63.6%) and trimethoprim-sulfamethoxazole (36.4%) and SP to penicillin (52.1%) and tetracycline (25.0%) were the highest. A variety of sequence types (STs) without a predominant type including numerous novel types were determined, especially for SP (39.6%). The spa typing provided a clonal assignment for all SAs (100%) and 24 SPs (50%) with three and two novel types, respectively. This study has provided an overview of SA and SP in the veterinary teaching hospital at UNZA. However, for a better understanding of these species regarding pathogenesis and transmission, further studies on the prevalence and characterization of SA and SP from veterinary staff, pet owners, and farm animals in Zambia is needed.     

Perspectives on canine and feline hepatozoonosis

Two species of Hepatozoon are currently known to infect dogs and cause distinct diseases. Hepatozoon canis prevalent in Africa, Asia, southern Europe, South America and recently shown to be present also in the USA causes infection mainly of hemolymphoid organs, whereas Hepatozoon americanum prevalent in the southeastern USA causes myositis and severe lameness. H. americanum is transmitted by ingestion of the Gulf Coast tick Amblyomma maculatum and also by predation on infected prey. H. canis is transmitted by Rhipicephalus sanguineus, in South America also by Amblyomma ovale, and has also been shown to be transmitted transplacentally. Hepatozoonosis of domestic cats has been described mostly from the same areas where canine infection is present and the exact identity of the species which infect cats, their pathogenicity and vectors have not been elucidated. The diagnosis of hepatozoonosis is made by observation of gamonts in blood smears, histopathology, PCR or serology. The main treatment for H. canis is with imidocarb dipropionate whereas H. americanum infection is treated with an initial combination of trimethoprim-sulfadiazine, pyrimethamine and clindamycin followed by maintenance with decoquinate. Treatment for both diseases has not been reported to facilitate complete parasite elimination and new effective drugs are needed for the management of these infections. Prevention of hepatozoonosis should be based on avoidance of oral ingestion of infected tick vectors and infected prey.     

Giardia duodenalis in small animals and their owners in Germany: A pilot study

Giardia duodenalis is a relevant gastrointestinal protozoan pathogen of humans and animals. This species complex consists of eight genetically different assemblages. Assemblages A and B are pathogenic to humans and pets, thus confer zoonotic potential. The risk of zoonotic transmission has been controversially discussed. The aim of this monocentric cross‐sectional pilot study was to investigate G. duodenalis assemblages in humans and pets living in common households in Berlin/Brandenburg (Germany). Samples from dogs, cats and humans sharing the same households were screened for Giardia infection by antigen‐detecting assays. All human samples were additionally analysed by a Giardia‐specific qPCR. Cyst quantification and sequences of different gene loci (triosephosphate isomerase (tpi), glutamate dehydrogenase (gdh), β‐giardin (bg) and for dogs SSUrDNA) were analysed. A total of 38 households (31 households with dogs and seven with cats) with 69 human individuals participated in the study. Initial antigen‐detecting assays revealed Giardia‐positive results for 13 (39%) canine, one (14%) feline and one human sample. Reanalysis of the human samples by qPCR revealed two more positive specimens (4%). Two of these three samples were identified as assemblage B at all tested loci. Success rate of assemblage typing for pet samples was generally low and comprised mainly the SSUrDNA locus only. Overall, six of 13 Giardia‐positive canine samples were typable (2× A, 1× co‐infection: A and B, 1× C; 2× D). One pair of samples (dog and human) from the same household had a similar but not identical assemblage B sequence at tpi locus. Assemblage A was also detected in the dog specimen, which hampered sequence analysis. In conclusion, although exhibiting limitations due to the sample size, our study highlights the need for better and standardized typing tools to distinguish G. duodenalis strains with higher resolution in order to perform proper case–control studies for a realistic estimation of zoonotic risk.     

Survey and molecular characterization of Cryptosporidium and Giardia spp. in owned companion animal, dogs and cats, in Japan

Compared with other countries, surveys of these parasites have been rarely performed in companion animals of Japan in spite of their significance for public health. Here, we investigated pet dogs and cats in Japan for the first time, and genetically analyzed the isolates to evaluate the risk of zoonotic infections. Seventy-seven fecal samples were collected from privately owned dogs and 55 samples from owned cats in Osaka city, Japan. Cryptosporidium oocysts were identified in 3/77 dogs (3.9%) and 7/55 cats (12.7%), and Giardia infection in 2/77 dogs (2.6%) and 1/55 cats (1.8%). Amplification of the target regions for genotyping was successful, Cryptosporidium isolates in dogs and cats were identified as C. canis and C. felis, respectively, and those of Giardia in dogs and cats were G. intestinalis Assemblages D and F. The discharge period of the oocysts varied within 3-16 weeks and that of the cysts was 12 weeks. To date, zoonotic types of both parasites have been identified in other animals in Japan, and further large-scale studies are needed to determine the distribution of zoonotic genotypes in these animals, especially those closely associated with humans.     

Old problems on a new playing field: Helminth zoonoses transmitted among dogs, wildlife, and people in a changing northern climate

Although surveillance is limited, indigenous residents at latitudes ranging from 53 to 73°N in Canada appear to have a higher occurrence of infection with some zoonotic parasites than the general population. Conversely, they are relatively naïve to other zoonotic parasites that have previously been unable to establish at northern latitudes. For those parasites that circulate among dogs, wildlife, and people, potential risk factors in the North include limited availability of veterinary services, presence of free-roaming dog populations, and consumption of locally harvested fish and wildlife. These regions are also experiencing some of the greatest impacts of climate change in North America, including increased temperature, precipitation, and frequency and severity of extreme weather. We review the current taxonomy, genetic diversity, host and geographic distributions, epidemiology and risk factors for 3 genera of helminths (Diphyllobothrium spp., Echinococcus spp., and Toxocara sp.) in Canada's North in order to identify climate-sensitive aspects of their ecology. Free-living stages of parasitic zoonoses endemic in the Arctic (such as Diphyllobothrium dendriticum, the cervid strain of Echinococcus granulosus, and Arctic strains of Echinococcus multilocularis) will experience trade-offs between enhanced survival under wetter conditions and increased mortality under warmer conditions. Climate change might also lead to the introduction and establishment in the Arctic of parasitic zoonoses previously restricted to the sub-Arctic, such as Diphyllobothrium latum, Toxocara canis, and the prairie strain of E. multilocularis. Molecular techniques applied in broad geographic surveys are needed to address critical knowledge gaps in the geographic distribution, genetic diversity, and public health significance of zoonotic helminths already in the circumpolar North, and to determine the current barriers to range expansion of temperate-adapted parasites into the North. Dogs will continue to play important roles in the North, including that of a “bridging” host between sylvatic cycles and human communities. In a warming north, increased opportunities for business, agriculture, and tourism favor importation of dogs and their parasites into a newly suitable environment. Collaborations among veterinarians, public health personnel, and policy-makers are needed to enhance surveillance and mitigate for dog-transmitted parasitic zoonoses in a changing North.     

Zoonotic enteric protozoa

A growing number of enteric protozoan species are considered to have zoonotic potential. Their clinical impact varies and in many cases is poorly defined. Similarly, the epidemiology of infections, particularly the role of non-human hosts, requires further study. In this review, new information on the life cycles and transmission of Giardia, Cryptosporidium, Entamoeba, Blastocystis and Balantidium are examined in the context of zoonotic potential, as well as polyparasitism and clinical significance.     

Novel hemotropic Mycoplasma species in white-tailed deer (Odocoileus virginianus)

Globally, hemotropic Mycoplasma spp. are emerging or re-emerging zoonotic pathogens that affect livestock, wildlife, companion animals, and humans, potentially causing serious and economically important disease problems. Little is known about hemotropic Mycoplasma spp. prevalence, host-specificity, or route of transmission in most species, including wildlife. DNA amplification by PCR targeting the 16SrRNA and the RNaseP genes was used to establish the presence and prevalence of hemotropic Mycoplasma spp. in a white-tailed deer (O. virginianus) population in eastern North Carolina. Sixty-five deer (89%) tested positive for hemotropic Mycoplasma spp. where sequence analysis of the 16SsRNA and the RNaseP genes indicated the presence of at least three distinct species. This study represents the first detection of three distinct hemotropic Mycoplasma species in white-tailed deer and the first report of two novel hemotropic Mycoplasma species.     

Baylisascariosis—Infections of animals and humans with ‘unusual’ roundworms

The nematode genus Baylisascaris (order Ascaridida, superfamily Ascaridoidea) contains nine relatively host-specific, parasite species of carnivores, omnivores, herbivores, carnivorous marsupials or rodents. They have a facultative heteroxenous life cycle, at least under experimental conditions. Eggs passed in faeces embryonate in the environment and the second-stage larva infective for both definitive and intermediate hosts develops. In intermediate hosts larvae migrate extensively through tissues, where they grow and moult to the third-stage, causing extensive damage. All Baylisascaris spp. are considered a potential cause of visceral, ocular and/or neural larval migrans in mammals including humans and in birds. This paper summarises our current knowledge on the prevalence, biology, pathogenicity and zoonotic significance of three Baylisascaris species: B. transfuga, B. schroederi and B. procyonis which have as definitive hosts bears, giant pandas and raccoons (occasionally dogs), respectively.     

Prevalence,species distribution and antimicrobial resistance patterns of methicillin-resistant staphylococci in Lithuanian pet animals

Background

The bacterial genus Staphylococcus consists of many species that causes infections in pet animals. Antimicrobial resistant staphylococci cause infections that are difficult to treat and they are important from the point of one health perspective. The aim of this study was to determine the prevalence of methicillin-resistant Staphylococcus (MRS) species, including methicillin-resistant S. aureus (MRSA) in diseased pet animals (Group A) and kennel dogs (Group B) in Lithuania and to characterize the isolates according to their antimicrobial resistance.

Results

Twenty-one MRS isolates were obtained from 395 clinical samples (5.3 %; CI 95 % 3.5-8.0) of Group A animals. Sixteen, four and one isolates were from dogs, cats and a pet rabbit, respectively. The mecA gene was present in 20 isolates, whereas one isolate was positive for the mecC gene. Twenty-one MRS isolates (20.0 %; CI 95 % 13.5-28.6) were obtained from the vagina of female dogs (n = 105) (Group B). All isolates carried the mecA gene. Twelve MRS species were isolated of which S. pseudintermedius was the most common (18/42) followed by S. haemolyticus (8/42) and S. lentus (4/42). MRSA was not found. All MRS strains were susceptible to vancomycin, linezolid, daptomycin and quinupristin/dalfopristin. Resistance to tetracycline (16/21), clindamycin (15/21) and erythromycin (14/21) was the most common types of resistance in Group A animals. Three isolates also demonstrated resistance to rifampin. Resistance toward gentamicin (16/21), ciprofloxacin (15/21), macrolides (15/21) and tetracycline (12/21) was the most common in kennel dogs (Group B). The most common genes encoding resistance to antimicrobials (excluding beta-lactams) in isolates from Group A pets were tetK (21/42), aph(3′)-IIIa (11/42) and aac(6'')-Ie-aph(2'''')-Ia (9/42).

Conclusions

A wide range of MRS species were found in pet animals in Lithuania. MRSA was not found.