Dog Fecal Samples Research Articles

Amphimeriasis in Ecuador-Prevalence, Knowledge, and Socio-Cultural Practices Among Indigenous Chachi and Montubios Populations: A Mixed-Methods Cross-Sectional Study.

Amphimerus, a liver fluke, is the causative agent of amphimeriasis, a foodborne disease acquired thought the consumption of infected raw or undercooked river fish-a practice embedded in traditional culinary customs. Amphimeriasis represents a significant public health issue and has been endemic in Ecuador since 2011, particularly among the Chachi Amerindians and Montubios populations residing in tropical ecoregions. By employing a mixed-methods research design, we conducted a community-based, cross-sectional study. A survey comprising of 63 questions on KAP was administrated in person to community members, health personnel, and academic staff in the two populations. Additionally, 67 semi-structured interviews were performed. Microscopy was achieved on 273 human and 80 dog fecal samples to detect Amphimerus eggs. A total of 86 questionnaires (54 Chachi) and 67 interviews (44 Chachi), out of 300 residents, were completed. Among the respondents, 31.4% were aware of Amphimerus, locally referred to as "liver worm". Although 79.1% reported not consuming raw fish, most admitted eating raw fish with lime juice and salt, a preparation known as "curtido", and 59.3% reported consuming smoked fish. Here, 86.1% of participants considered "liver worm" a serious disease, and 55.8% recognized raw or marinated fish as a potential transmission route. The Chachi showed a preference for smoked fish, whereas the Montubios favoured "curtido". The prevalence of Amphimerus infection was 23% in humans and 16.2% in dogs. Differences in KAP were observed between infected and non-infected individuals. Local health and academic personnel demonstrated insufficient knowledge about amphimeriasis. Some religious individuals refrained from participating, stating that they were "with God". Despite the high prevalence of Amphimerus infection in both humans and dogs, knowledge about the parasite, the disease, and its transmission routes remains limited. Health education initiatives should be designed to modify the population's KAP. It is crucial for national and local health authorities, as well as religious leaders, to be informed and actively involved in the prevention and control of amphimeriasis.

Emerging Echinococcus tapeworms: fecal PCR detection of Echinococcus multilocularis in 26 dogs from the United States and Canada (2022-2024).

To report quantitative PCR (qPCR) detection of Echinococcus multilocularis DNA in fecal samples from 26 dogs in the US and Canada. 26 dogs with fecal samples submitted for parasite screening by qPCR. Dog signalment, presenting concern, preventive care, and outcomes were obtained from the primary veterinarian via email or telephone, where available. Echinococcus multilocularis was detected in 26 of 2,333,797 dog fecal samples by reference laboratory fecal qPCR surveillance between March 2022 and July 2024. Seventeen E multilocularis-detected samples were sequenced as European haplotypes (E3/E4). Taenia-type eggs were identified by zinc sulfate centrifugal flotation in 8 of 17 samples (47%). Dogs were from the US (n = 16) and Canada (10). Ten dogs had gastrointestinal signs (diarrhea) reported on initial presentation. Clinical history revealed that some dogs were receiving a monthly antiparasitic preventive in the 6-month period prior to sampling (n = 10) and had regular wildlife (rodent) exposure (13). Twenty-five dogs were subsequently confirmed to have received treatment with praziquantel for detected E multilocularis, and 25 of these dogs were qPCR negative 3 to 5 weeks after treatment. Veterinary awareness of endemic risk regions for E multilocularis and its emergence in novel areas (Colorado, Nevada, Wyoming, Montana, Illinois, Washington, Idaho, Kansas, and Oregon) are key for One Health. Dogs can serve as sentinels for Echinococcus tapeworm risk, and detection of E multilocularis tapeworms in dogs through routine qPCR fecal screening can alert clinicians to zoonotic concern and common environmental exposure risk.

A pilot study on the epidemiology, diagnosis and characterization of Echinococcus granulosus sensu lato in sheep, goats and dogs in Siirt province of Türkiye revealed remarkable adaptation of Echinococcus canadensis (G6/G7) in goats

Cystic echinococcosis (CE), caused by Echinococcus granulosus sensu lato (s.l.), represents one of the most significant zoonotic diseases globally, affecting both humans and animals. The objective of this study was to ascertain the prevalence of E. granulosus sensu lato in sheep and goats in a pilot region with a one-year slaughterhouse follow-up period and to determine the genetic differences and haplotypes among sheep, goat, and dog isolates. To this end, the prevalence of CE cysts was determined by monitoring the slaughter of sheep and goats at least three days a week at a slaughterhouse in the Siirt province of Türkiye during 2023. Additionally, faecal samples were collected from stray dogs and analysed using both flotation and molecular techniques. The presence of CE cysts was identified in 569 (11.12 %) of the 5119 sheep and 66 (2.31 %) of the 2860 goats after slaughtering. The highest positivity was observed in November (20.39 %), while the lowest was recorded in July (5.62 %). Of the sheep that detected positive, 25 (4.39 %) were less than one year old, while 544 (95.61 %) were older than one year. Of the infected sheep, 26 (4.57 %) were male and 543 (95.43 %) were female. 204 (35.85 %) sheep exhibited fluid-filled CE cysts, 338 (59.40 %) displayed calcification, and 27 (4.75 %) demonstrated the presence of newly developed cysts. The highest positivity was observed in December (5.83 %), while the lowest was recorded in May (0.62 %) in goats. Of the positive goats, two (3 %) were less than one year old, while the remaining 64 (97 %) were older than one year. Of the goats infected with CE cysts, 10 (15.15 %) were male and 56 (84.85 %) were female. Of the cysts, 56.1 % were fluid-filled, 42.4 % were calcified and 1.5 % were newly developed. Following DNA sequence analysis of CE cyst isolates obtained from the slaughterhouse, all 61 sheep sequences were identified as E. granulosus s.s. (G1/G3). Of the 13 goat isolates, seven were identified as E. granulosus s.s. (G1/G3), while the remaining six were classified as E. canadensis (G6/G7). The centrifugal flotation method was employed to detect the presence of Isospora spp. oocysts in eight dogs, Toxocara canis and hookworm eggs in three dogs each, and Dipyllidium caninum eggs in one dog. A total of 54 dog faeces were examined. No Taeniid eggs were observed in any of the dogs. Following PCR analysis of the mt-CO1 gene region in the dog faecal samples, four samples were positive for a 875 bp band. Only one of these bands was suitable for sequence analysis, which confirmed it as E. granulosus s.s. (G1/G3).

First Sero-Molecular Diagnosis of Toxoplasma gondii and Toxocara spp. Infections in the Police Dogs and Their Trainers in Iran.

Toxoplasma gondii (T. gondii) and Toxocara spp. are two types of parasites that can infect humans and various animals, including dogs. Police dogs and their trainers have a vital role in law enforcement, and their health and well-being are crucial for them to effectively carry out their duties. No study has yet been conducted on the prevalence of T. gondii and Toxocara spp. infections among police dogs and their trainers in Iran. The objective of this study was to determine the sero-molecular prevalence of T. gondii and Toxocara spp. infections in police dogs and their trainers in Tehran, the capital of Iran. In Tehran province, the anti-narcotics police have nearly 200 well-trained police dogs. Each dog is assigned a dedicated trainer and upon completing missions, is housed separately in a designated area. In the present study, a total of 150 samples were gathered. These included 50 blood samples from randomly selected police dogs, 50 fecal samples from the same dogs, and 50 blood samples from their trainers. The Modified Agglutination Test (MAT) was performed to detect T. gondii antibodies in dog blood samples and the ELISA system was utilized to identify anti-Toxoplasma and anti-Toxocara antibodies in the sera of the dog trainers. A specific segment of the SAG2 and ITS genes were amplified via nested-PCR in order to molecularly detect T. gondii in human blood samples and Toxocara spp. in dog fecal samples. Regarding serological findings, the prevalence of T. gondii in dog and human blood samples was 4% (2/50) and 10% (5/50), respectively. According to reports, the seroprevalence of Toxocara spp. in human blood samples was 6% (3/50). No statistically significant association was found between the prevalence of the examined parasites and variables (age, sex, and breed) in dogs, as well as the age variable in military personnel. Molecular findings showed that out of the 50 dog fecal samples and 50 human blood samples, there was no presence of Toxocara spp. and T. gondii, respectively. Understanding the prevalence of parasitic infections helps public health officials assess the risk to human and animal populations. This information can guide the development of prevention and control measures to reduce the spread of these infections. Overall, the prevalence of parasitic infections, particularly T. gondii and Toxocara spp., in police dogs and their trainers remains uncertain and necessitates further in-depth research.

Environmental contamination with feces of free-roaming dogs and the risk of transmission of Echinococcus and Taenia species in urban regions of southeastern Iran

BackgroundDogs are the most important definitive hosts of zoonotic taeniid helminths worldwide. Different Echinococcus and Taenia species of domestic and wild carnivores pose a potential risk to human population. High populations of free-roaming dogs (FRDs) in urban areas of Iran and widespread contamination of the environment with dog feces is a potential source of infecting people living in the urban regions with cystic echinococcosis (CE). Our knowledge on the risk of CE transmission in the urban settings in the endemic regions is limited. The present study surveyed the species and genotypes of E. granulosus sensu lato and other taeniids by examining feces of free-roaming dogs in the urban areas in the city of Kerman, southeastern Iran.MethodsThe city was divided into 100 consecutive blocks of which 25 blocks were randomly selected. Fecal samples of FRDs were counted, mapped and fresh samples were collected. Then Zinc chloride flotation, and sequential sieving was performed, and the samples were examined under an inverted microscope. Single individual taeniid eggs were isolated, partial nad1 gene was amplified and sequenced to identify species and genotypes.ResultsIn total 5607 fecal samples of dogs were mapped and 83 fresh samples were collected. Taeniid eggs were detected in nine fecal samples (10.8%) from seven out of the 25 city blocks (28.0%). Echinococcus eggs were found in four samples (4.8%) from three city blocks, two samples containing E. granulosus sensu stricto (2.4%), two samples containing E. canadensis G6/7 (2.4%). In addition, three samples contained eggs of Taenia hydatigena (3.6%), and one sample of Taenia serialis (1.2%).ConclusionsThis study documented the potential risk of CE transmission to humans resulting from the feces of dogs roaming freely in urban areas.Graphical

Optimization of CRISPR/Cas12a detection assay and its application in the detection of Echinococcus granulosus

Cystic echinococcosis, resulting from infection with Echinococcus granulosus, poses a significant challenge as a neglected tropical disease owing to the lack of any known effective treatment. Primarily affecting under-resourced, remote, and conflict-ridden regions, the disease is compounded by the limitations of current detection techniques, such as microscopy, physical imaging, ELISA, and qPCR, which are unsuitable for application in these areas. The emergence of CRISPR/Cas12a as a promising tool for nucleic acid detection, characterized by its unparalleled specificity, heightened sensitivity, and rapid detection time, offers a potential solution. In this study, we present a one-pot CRISPR/Cas12a detection method for E. granulosus (genotype G1, sheep strain) integrating recombinase polymerase amplification (RPA) with suboptimal protospacer adjacent motif (PAM) and structured CRISPR RNA (crRNA) to enhance reaction efficiency. The evaluation of the assay's performance using hydatid cyst spiked dog feces and the examination of 62 dog fecal samples collected from various regions of Western China demonstrate its efficacy. The assay permits visual observation of test results about 15 minutes under blue light and displays superior portability and reaction speed relative to qPCR, achieving a sensitivity level of 10 copies of standard plasmids of the target gene. Analytic specificity was verified against four tapeworm species (E. multilocularis, H. taeniaeformis, M. benedeni, and D. caninum) and two other helminths (T. canis and F. hepatica), with negative results also noted for Mesocestoides sp. This study presents a rapid, sensitive, and time-efficient DNA detection method for E. granulosus of hydatid cyst spiked and clinical dog feces, potential serving as an alternative tool for field detection. This novel assay is primarily used to diagnose the definitive host of E. granulosus. Further validation using a larger set of clinical fecal samples is warranted, along with additional exploration of more effective approaches for nucleic acid release.

Detection and characterization of canine corona virus infection: an enteric approach

Canine corona virus (CCoV) infection is a new emerging disease affecting dogs. It results in low mortality but considerable morbidity in dogs with intermediate to moderate enteritis. In India, serological research on dogs has revealed the existence of antibodies, demonstrating the prevalence of the condition among canines. The aim of the present study was to investigate the prevalence of CCoV infection in dogs suffering from gastroenteritis, and genetic analysis of circulating virus strains. The current study was designed to test fecal samples of dogs with gastroenteritis suspected for the presence of CCoV. Using primers based on the M-gene, PCR was used to amplify the RNA (c-DNA) templates retrieved from the fecal samples. RT-PCR based detection of CCoV showed positivity of 30% (15/50). The phylogentic analysis revealed that two samples had nt/aa identity levels of 100%/100% to the CCoV isolate from China and 99.19/100% of nt/aa identity with that from Korea. One of the samples showed divergence and revealed 95.63%/94.37% nt/aa identity with the China isolate and 95.15%/97.06% nt/aa identity with Indian isolates. The recombination analysis indicated a mutation rate in one of the samples. In brief, the present study illustrates the circulation of canine corona virus in dogs suffering from gastroenteritis. The present findings emphasize the need for vaccination of dogs for prevention, as a high prevalence of viral gastroenteritis was found in these dogs.

Comparison of different molecular protocols for the detection of Uncinaria stenocephala infection in dogs

The present study aims to assess the performance of different molecular targets using various matrices of samples for the detection of Uncinaria stenocephala (US) in hookworm infected dogs. To this end, the DNA extraction was performed on the following matrices of samples: (i) larvae of US obtained from experimentally infected dogs with US with different larvae counts per microliter (µl); (ii) pure US eggs suspension in distilled water with different egg counts per µl; (iii) spiked dog fecal samples with different US eggs per gram (EPG) of feces; (iv) feces from dogs naturally infected with hookworm eggs; (v) fecal suspension with hookworm eggs recovered from the FLOTAC apparatus. All the samples were tested with four different PCR protocols targeting specific regions for the detection of both hookworms US and AC as follows: Protocol A (ITS1, 5.8 S, ITS2) and Protocol B (18 S) for the detection of both species, Protocol C (ITS1) for the detection of AC and Protocol D (ITS1) for the detection of US. The best results were obtained with DNA extracted from US larvae matrix obtained from experimentally infected dogs, showing a detection limit of 3.5 larvae/ml for the protocols A, B and D. A moderate correlation was found between the FLOTAC technique and PCR protocols B and D with respect to fecal samples from dogs naturally infected with hookworms. Indeed, PCR protocols B (18 S) and D (ITS1) gave the best results for feces and fecal suspension from naturally infected dogs. However, all the PCR protocols used showed lower sensitivity than FLOTAC technique. Perhaps, isolating US eggs in advance could help to obtain better quality and quantity of DNA, avoiding some notable factors such as inhibitors present in faecal samples. However, a further study is needed to evaluate and standardise a protocol for the recovery of parasitic elements, that could be applied prior to DNA extraction. Therefore, this could lead to a better amplification of US eggs DNA. In conclusion, our results showed that the type of sample (sample-matrix) used for the DNA extraction samples is crucial, as this affects the diagnostic sensitivity of the technique.

Isolation of multi-drug-resistant strains of Escherichia coli from faecal samples of dogs and cats from Harare, Zimbabwe.

The escalation of antimicrobial resistance (AMR) in recent years has been of major public health concern globally. Escherichia coli are amongst the bacteria that have been targeted for AMR surveillance due to their ability to cause infection in both animals and humans. Their propensity to produce extended spectrum beta-lactamases further complicates the choices of treatment regimens. To investigate the prevalence of antimicrobial-resistance in E. coli strains isolated from faecal samples of dogs and cats from selected veterinary surgeries and animal shelters from Harare, Zimbabwe. A cross-sectional study was carried out to select animals by a systematic random procedure. Faecal samples were collected for culture and isolation of E. coli. Their susceptibility to antimicrobial drugs was assessed using the disc diffusion method. A total of 95% (133/140) of the samples from cats (n=40) and dogs (n=93) yielded E. coli. Resistance was recorded for ampicillin (45.9%), trimethoprim-sulphamethoxazole (44.4%), nalidixic acid (29.3%), ceftazidime (15.8%) and azithromycin (12.8%), but not for gentamicin and imipenem. A total of 18% of the isolates were multi-drug-resistant where resistance to nalidixic acid, ampicillin and trimethoprim-sulphamethoxazole predominated. We observed relatively high AMR of E. coli strains against ampicillin. The isolation of multi-drug-resistant strains of E. coli may signal the dissemination of resistance genes in the ecosystem of these bacteria which may have a public health impact.

Rapid Discriminative Identification of the Two Predominant Echinococcus Species from Canine Fecal Samples in the Tibetan Region of China by Loop-Mediated Isothermal Amplification-Lateral Flow Dipstick Assay.

Echinococcosis poses a significant concern in the fields of public health and veterinary care as it can be transmitted between animals and humans. The primary endemic subtypes are cystic echinococcosis (CE) and alveolar echinococcosis (AE), which result from infestation by Echinococcus granulosus and Echinococcus multilocularis, respectively. A prominent epidemic of echinococcosis greatly affects the Tibet Autonomous Region (TAR) in China. A new technique called the loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) test is introduced in this research to differentiate between E. granulosus and E. multilocularis using their repetitive genetic sequences. The test is characterized by its portable nature, simple operation, quick result production, high sensitivity, and low susceptibility to aerosol contamination. The LAMP-LFD method demonstrated an exceptional minimal detection limit, reaching levels as low as approximately 1 fg/μL (femtogram per microliter) of genomic DNA. The assay's specificity was assessed, and no cross-reactivity was seen. A total of 982 dog fecal samples were collected from 54 counties in the TAR region between July 2021 and June 2022. The established method underwent validation using a commercially available ELISA kit. The agreement rate between the LAMP-LFD and ELISA methods was 97.25%, with a sensitivity of 96.05% and a specificity of 97.35%. The assay described in this study improves specificity by using a double-labeled probe, and it reduces the risk of false-positive results caused by aerosol contamination through the use of a sealed device. This makes it a suitable choice for quickly and accurately identifying the two main types of Echinococcus in field settings.

Management to Reduce Impact of Cerebral Coenurosis in Bonga Sheep in Kafa Zone, South Western Ethiopia

Coenurosis, a parasitic and fatal disease, is widespread in the highlands of Ethiopia where 75% of the country’s sheep population are found. There is significant prevalence of the disease in the current study area. Therefore, the current study was conducted with the objectives to reduce impact of cerebral coenurosis in sheep production in the study area through integrating different prevention and control options and to raise the community awareness about disease. Questionnaire surveys, coproscopic examination of dog faeces and household level surveillance was used to collect data on reported coenurosis cases and taeniid infections. Awareness creation trainings and regular deworming of dogs were used as an intervention to reduce the prevalence of the disease. A total of 107, 134, 153, and 124 dogs were dewormed during the first, second, third, and fourth rounds during the study period respectively. Eggs were detected in 58.53% (95% CI: 47.4- 68.86) of pre-deworming fecal samples of dogs. Eggs were detected in 24.18% (95% CI: 40.0–49.7) of post-intervention fecal samples, 34.35% less than pre-intervention. At the beginning of the intervention the level of awareness of the farmers in the intervention sites about the cause of the disease was 12.2% which was increased to 51.03% at the end of the intervention. The community-based integrated animal health management approach for the control of coenurosis has the potential to be scaled-up across the country, thereby reducing economic losses of communities where the disease is endemic.

Parasitological investigation of soil biotope in the territory of Moscow

The purpose of the research is to conduct a sanitary and parasitological examination of soil biotopes on the territory of Moscow. In addition to traditional soil studies, we conducted coproscopic analyzes of samples of dog feces collected from the soil surface in Spring.Materials and methods. 83 combined soil samples were taken in 9 administrative districts of Moscow using the envelope method at a depth of up to 10 cm. From each combined sample, 4 samples were taken and examined using the Romanenko method according to MUK 4.2.2661-10 “Methods of sanitary and parasitological research”. From the same areas where soil was collected, 365 dog fecal samples were collected. Feces were studied using a combined flotation method. Soil and fecal samples were taken in Spring of 2023 during a period of positive temperatures immediately after the snow melted. Microscopy was performed using a Motic BA410T microscope. Identification of the detected objects was carried out on the basis of morphometric data. Species differentiation of Toxocara canis and T. cati eggs was carried out based on differences in the size of the eggs and the structure of the outer shell.Results and discussion. The city's soil is contaminated with pathogens of parasitic diseases in 9.3% of samples. Toxocara eggs were found (5.7%), of which T. canis in 3.9% of samples, and T. cati in 1.8% of samples. Eggs of Capillaria sp. detected in 1.5%, eggs of Trichuris sp. at 0.9%. Eggs of Hymenolepis sp. and coccidia were detected in 0.6% of samples. In dog feces, pathogens of parasitic diseases were detected in 3.3% of samples. Toxocara sp. eggs were found in 1.4% of samples, isospores in 0.8%, sarcocystis in 0.8%, Toxascaris leonina eggs in 0.3%. Toxocara sp. eggs are the leaders in the frequency of detection in the soil; these are mainly viable eggs with larvae. This is consistent with the data that T. canis eggs are most often recorded in anonymized dog feces. Eggs of Trichuris sp. and Capillaria sp. found in soil with formed viable larvae. The findings show the potential of urban soils as a parasite transmission factor for humans and susceptible animals, and dogs play a leading role in the contamination of urban soils with Toxocara sp. eggs.

Investigation of the distribution of some Gastrointestinal Helminths and protozoa in free-ranging dogs in Karbala province

This study aimed to determine the prevalence of intestinal parasites in stray dogs in urban and rural areas. From October 2022 to March 2023 80 fecal samples, which examined by using simple zinc sulfate flotation and Lugol’s solution staining then examined by macroscopically and microscopically for the presence of worm eggs and protozoal oocysts. The overall prevalence of infected dogs in this study is to identify the gastrointestinal parasites and Protozoa in stray dogs. The current study includes seven genera of intestinal helminths and protozoa as follows: three cestodes (Taenia sp. 12%), four nematodes (Toxocara canis 56%, Toxoascara canis. 16%) and two protozoa (Giardia sp. 20% and Cryptosporidium sp. 8%) from all fecal samples of dogs were collected from different regions of Kerbala. The results of high rates of infection were during winter months. The high prevalence of intestinal helminths in the dog’s population suggests the need for more efficient control measures. The high predominance of T. canis, and Giardia spp. suggested that dogs could play an active role in the transmission of zoonotic parasites in this area of Iraq. Educating the dog hunters and increasing their health awareness should be considered in the control program.
 The results of the present study provide relevant ‘‘baseline’’ data for assessing the effectiveness of future control strategies against canine parasitic infections.

Gastrointestinal parasites of dogs and foxes in the Zanjan province of Iran: With an emphasis on Echinococcus species

Canids harbor many zoonotic parasites and play an important role in the spread of parasites in the human environment. Estimation of parasitic infection among canids as definitive hosts may help competent authorities design efficient control programs. This study was conducted to determine the prevalence of intestinal parasites in dogs and foxes with an emphasis on Echinococcus spp. A total of 500 fecal samples of dogs and 30 fecal samples of foxes were studied in the summer, autumn, and winter of 2021 in the Zanjan province using the formalin-ethyl acetate concentration technique, followed by multiplex PCR. The overall prevalence of gastrointestinal parasite infection was estimated to be 19.05%. The prevalence was 24.8%, 10.2%, and 26.7% in stray, shelter dogs and foxes, respectively. No parasites were found among pet and guard dog samples. PCR results on Taenidae eggs showed that 2.4% of samples were positive for Echinococcus granulosus and none contained E. multilocularis. Noteworthy is that E. granulosus was identified only in stray dog samples. The higher prevalence of E. granulosus infection in stray dogs in this province emphasizes the importance of monitoring the food sources consumed by these animals.

Outcome of parasitological examinations in dogs in Germany: a retrospective survey

Dog faecal samples examined from January 2019 to December 2019 were retrospectively analysed for frequency of endoparasites. The examinations were performed with several different methods: 29,219 samples were examined by flotation method and sodium acetate-acetic acid-formalin concentration (SAFC) technique, 1,330 samples by Baermann-Wetzel migration technique, 12,221 samples using a Giardia coproantigen enzyme-linked-immunosorbent assay (ELISA), 1,180 samples using a Cryptosporidium coproantigen ELISA, 1,671 samples by polymerase chain reaction (PCR) testing for Giardia duodenalis and 447 samples by PCR testing for Cryptosporidium spp.. A total of 7.1% of the samples were positive for parasites in the microscopical examination using the flotation method and SAFC technique. The parasites found included Cystoisospora spp. (2.8%), Giardia duodenalis (2.3%), Ancylostomatidae (1.8%), Toxocara canis (1.6%), Trichuris vulpis (0.7%), Toxascaris leonina (0.5%), Capillaria spp. (0.2%), Angiostrongylus vasorum (0.2%), Crenosoma vulpis (0.1%), Taeniidae (0.1%), Sarcocystis spp. (0.03%), Dipylidium caninum (0.01%), Diphyllobothrium latum (< 0.01%), Spirurida (< 0.01%) and Opisthorchiidae (< 0.01%). Using the Baermann-Wetzel migration technique, Angiostrongylus vasorum was found in 0.75% and Crenosoma vulpis in 0.3% of the samples. ELISAs for Giardia duodenalis and Cryptosporidium spp. revealed 13.9% and 1.0% positive faecal samples, and Giardia duodenalis and Cryptosporidium spp. PCRs 19.4% and 2.0%, respectively. Dogs in the first year of life were more frequently infected with parasites than older animals. In the microscopic examination using flotation method and SAFC technique, the significantly highest detection rates were found in dogs up to six months of age (p < 0.001).

In Vitro Assessment of Postbiotic and Probiotic Commercial Dietary Supplements Recommended for Counteracting Intestinal Dysbiosis in Dogs.

Many environmental aspects influence the preservation of a beneficial microbiome in dogs, and gut dysbiosis occurs when imbalances in the intestinal ecosystem cause functional changes in the microbial populations. The authors evaluated the effects of two specific commercial dietary supplements: a combination of a postbiotic and prebiotics (Microbiotal cane®) and a probiotic product (NBF 1®) recommended for counteracting intestinal dysbiosis in dogs, on the gut canine microbiota composition and its metabolic activities (production of short-chain fatty acids). The investigation was performed using an in vitro fermentation system inoculated with dog fecal samples. Microbiotal cane® promoted a more immediate increase in Lactobacillus spp. after the first 6 h of fermentation, whereas NBF 1® promoted the increase at the end of the process only. The two supplements supported an increase in the Bifidobacterium spp. counts only after 24 h. The in vitro abilities of Microbiotal cane® and NBF 1® to increase selectively beneficial bacterial groups producing acetic, propionic, and butyric acids suggest a possible positive effect on the canine gut microbiota, even if further in vivo studies are needed to confirm the beneficial effects on the intestinal health.

Prevalence and first molecular identification of Sarcocystis species in feces of domestic dogs (Canis familiaris) in Egypt

BackgroundSarcocystis species are obligatorily heteroxenous protozoan parasites with predator–prey life cycles. Global Knowledge about the epidemiology and the distribution pattern of different Sarcocystis species in dog feces are very scarce. Therefore, the current investigation was conducted to declare the occurrence of Sarcocystis in the fecal specimens of the most common canids in Egypt, the domestic dogs, and to identify the species present using various parasitological and molecular approaches.MethodsA total of 100 dog fecal samples were collected and screened using fecal sugar flotation test for the presence of Sarcocystis oocysts/sporocysts. Additionally, thirty samples were used for genomic DNA extraction. The 18S rRNA gene fragment was the target of primers for a PCR, followed by purification and sequencing of the amplicons.ResultsCurrently, the results obtained reviewed that 4% of fecal samples were positive for Sarcocystis spp. using LM. Additionally, Sarcocystis spp. were verified in sixteen dogs (53.3%, 16/30) using PCR and subsequent sequencing protocols. Statistically, insignificant difference in prevalence of sarcocystosis relative to age and gender was noticed. Morphologically, the detected sporocysts measured 13.2–16.0 × 9.4–11 μm. Based on the 18S rRNA gene, sequencing analysis of amplicons from sporocysts DNA revealed 99.82% nucleotide homology with published S. tenella partial nucleotide sequences from sheep in Iraq and Iran.ConclusionsThis is the first molecular evidence in support of the final host role of domestic dogs in the life cycle of S. tenella in Egypt, which provides a precious diagnostic tool for further epidemiological studies and for the assessment of the effectiveness of control measures for this disease.

Validation of method for faecal sampling in cats and dogs for faecal microbiome analysis

BackgroundReproducible and reliable studies of cat and dog faecal microbiomes are dependent on many methodology-based variables including how the faecal stools are sampled and stored prior to processing. The current study aimed to establish an appropriate method for sampling and storing faecal stools from cats and dogs which may also be applied to privately-owned pets. The approach investigated the effects of storing faeces for up to 12 h at room temperature and sampling from various locations within the stool in terms of microbial diversity, relative taxa abundances and DNA yield. Faeces were collected from 10 healthy cats and 10 healthy dogs and stored at room temperature (20 °C). Samples were taken from various locations within the stool (the first emitted part (i), the middle (ii) and the last emitted end (iii), at either surface or core) at 0, 0.5, 1, 2, 3, 6 and 12 h, stabilised and stored at -80 °C. DNA was extracted from all samples, using Illumina NovaSeq.ResultsFaecal bacterial composition of dogs and cats shown no statistically significant differences in alpha diversity. Bacteroidetes, Firmicutes, Proteobacteria and Actinobacteria were the most prevalent phyla. Cat and dog samples were characterized by a dominance of Prevotella, and a lack of Fusobacterium in feline stools. Room temperature storage of cat and dog faecal samples generally had no significant effect on alpha diversity, relative taxa abundance or DNA yield for up to 12 h. Sampling from regions i, ii or iii of the stool at the surface or core did not significantly influence the outcome. However, surface cat faecal samples stored at room temperature for 12 h showed a significant increase in two measures of alpha diversity and there was a tendency for a similar effect in dogs. When comparing samples with beta diversity measures, it appeared that for dog and cat samples, individual effect has the strongest impact on the observed microbial diversity (R2 0.64 and 0.88), whereas sampling time, depth and horizontal locations significantly affected the microbial diversity but with less impact.ConclusionCat and dog faeces were stable at room temperature for up to 12 h, with no significant changes in alpha diversity, relative taxa abundance and DNA concentration. Beta diversity analysis demonstrated that despite an impact of the sampling storing time and the surface of the sampling, we preserved the identity of the microbial structure linked to the individual. Finally, the data suggest that faecal stools stored for > 6 h at room temperature should be sampled at the core, not the surface.

Development of an LFD-RPA Assay for Rapid Detection of Pentatrichomonas hominis Infection in Dogs.

Pentatrichomonas hominis is a trichomonad protozoan that infects the cecum and colon of humans and other mammals. It is a zoonotic pathogen that causes diarrhea in both animals and humans. As companion animals, dogs infected with P. hominis pose a risk of transmitting it to humans. Current methods, such as direct smears and polymerase chain reaction (PCR), used for P. hominis detection have limitations, including low detection rates and the need for specialized equipment. Therefore, there is an urgent need to develop rapid, sensitive, and simple detection methods for clinical application. Recombinase polymerase amplification (RPA) has emerged as a technology for rapid pathogen detection. In this study, we developed a lateral flow dipstick (LFD)-RPA method based on the highly conserved SPO11-1 gene for detecting P. hominis infection by optimizing the primers, probes, and reaction conditions, and evaluating cross-reactivity with genomes of Giardia duodenalis and other parasites. The LFD-RPA method was then used to test 128 dog fecal samples collected from Changchun. The results confirmed the high specificity of the method with no cross-reactivity with the five other parasites. The lowest detection limit of the method was 102 copies/µL, and its sensitivity was 100 times higher than that of the conventional PCR method. Consistent with the positivity rate observed using nested PCR, 12 samples (out of 128) tested positive using this method (positivity rate, 9.38%). In conclusion, the LFD-RPA method developed in this study represents a simple and sensitive assay that allows for the rapid detection of P. hominis infection in dogs, especially in this field.

Giardiasis and diarrhea in dogs: Does the microbiome matter?

Giardia duodenalis (Gd) causes intestinal parasitosis. The involvement of the intestinal microbiome in determining the infection's clinical phenotype is unknown. Investigate the fecal microbiome features in dogs with giardiasis. Cross-sectional study, including fecal samples of kenneled dogs with Gd diagnosed by fecal Giardia antigen dot ELISA. The fecal microbial compositional characteristics and dysbiosis index (DI) were compared between diarrheic and nondiarrheic dogs. Fecal samples of 38 Gd-infected dogs (diarrheic, 21; nondiarrheic, 17) were included. No differences were found in Faith's phylogenic diversity and beta diversity (weighted UniFrac distances) and in specific taxa abundances at the phylum, genus, and species levels, as well as in alpha and beta diversities between diarrheic and nondiarrheic dogs, and also when divided by sex or age. Among diarrheic dogs, alpha diversity was higher in males than in females (pairwise Kruskal-Wallis, q = 0.01). Among males, fecal abundances of the genus Clostridium (W = 19) and Clostridium spiroforme species (W = 33) were higher in diarrheic compared to nondiarrheic dogs. In diarrheic dog fecal samples, Proteobacteria were more prevalent (W = 1), whereas Verrucomicrobia were less prevalent in dogs <1 year of age than in older dogs. The fecal sample DI of 19 diarrheic and 19 nondiarrheic dogs was similar (median, -0.2; range, -4.3 to 4.5 and median, -1.0; range, -4.3 to 5.8, respectively). The fecal microbial composition of symptomatic and asymptomatic dogs with giardiasis is similar. Based on fecal DI, giardiasis is not characterized by prominent dysbiosis. Other host and parasite characteristics might determine the severity of giardiasis in dogs.