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1.
Arq. bras. med. vet. zootec. (Online) ; 73(2): 522-524, Mar.-Apr. 2021.
Article in English | LILACS, VETINDEX | ID: biblio-1248921

ABSTRACT

Trichostrongylid em ovelhas foi estudado no Distrito Zhob, Balochistan. Foram coletados 120 tratos gastrointestinais (GIT) de ovelhas do matadouro do distrito. Estas amostras foram processadas para isolamento e identificação de nematódeos tricostrônquicos no Laboratório do Hospital Veterinário do Distrito Zhob. A taxa de prevalência geral foi de 39,1% em machos e 60,8% em fêmeas (p=0,001). A taxa de prevalência em duas raças viz Balochi e Rakhshani foi de 58,3% e 41,6%, respectivamente (p=0,01). A prevalência da espécie observada com Trichostrongylus foi 19,1%, Haemonchus foi 20,8%, Cooperia foi 29,1% e Nematodirus foi 30,8% (p=0,087). Quanto a quantidade de espécies infestadas pelas ovelhas, um único tipo de parasita estava em 32,5% de animais, dois tipos de espécies parasitárias em 36,3% dos animais e três tipos de espécies parasitárias em 30,8% dos animais (p=0,366).(AU)


Subject(s)
Animals , Trichostrongylosis/veterinary , Trichostrongylosis/epidemiology , Trichostrongylus/isolation & purification , Sheep/parasitology , Nematodirus/isolation & purification , Haemonchus/isolation & purification , Pakistan , Gastrointestinal Tract/parasitology
2.
Vet Parasitol Reg Stud Reports ; 22: 100474, 2020 12.
Article in English | MEDLINE | ID: mdl-33308728

ABSTRACT

Two methodologies were tested to isolate pure Trichostrongylus colubriformis strains from naturally infected sheep. Also, the in vitro susceptibility status to commercial anthelmintic (AH) drugs was determined in these strains. These methods were performed in a tropical region of Mexico where naturally infected sheep and goats host Haemonchus contortus, T. colubriformis and Oesophagostomum columbianum. For the first strain, a group of 6 naturally infected lambs from the "Paraiso" farm were treated with closantel (subcutaneous (SC), 10 mg/kg). On day 10 post-treatment, the eggs per gram (EPG) of faeces were determined with the McMaster technique. The faeces from the two lambs with the highest EPG were used for coprocultures (4 days, 28 °C). The L3 larvae were recovered and identified as T. colubriformis (69%) and O. columbianum (31%). The latter was removed by 10-day refrigeration (4-5 °C) followed by sieving (25 µm), resulting in 100% T. colubriformis (PARAISO strain). The second strain was isolated using repeated doses of levamisole (LEV, SC 7.5 mg/kg) in an 8-year-old sheep. The sheep had 1700 EPG before the LEV treatments and maintained 1300 EPG after both LEV treatments (day 10). The coproculture (4 days, 28 °C) after the second treatment contained 100% T. colubriformis (FMVZ-UADY strain). The in vitro AH susceptibility was determined using the egg hatch test for benzimidazole (BZ), and the larval migration inhibition test for ivermectin (IVM) and LEV. The PARAISO strain was BZ- and LEV-susceptible, and IVM-resistant. Meanwhile, the FMVZ-UADY strain was BZ- and IVM-susceptible, and LEV-resistant. The conditions where these two protocols could be used in other parts of the world were discussed.


Subject(s)
Sheep Diseases , Sheep/parasitology , Trichostrongylosis/veterinary , Trichostrongylus/isolation & purification , Animals , Mexico , Parasite Egg Count/veterinary , Sheep Diseases/drug therapy , Sheep Diseases/parasitology , Trichostrongylosis/drug therapy
3.
Parasit Vectors ; 13(1): 553, 2020 Nov 07.
Article in English | MEDLINE | ID: mdl-33160415

ABSTRACT

BACKGROUND: Trichostrongylus is one of the most important zoonotic trichostrongylid nematodes, infecting mostly livestock. Data on its genetic characteristics are lacking in Iran. METHODS: We determined the phylogenetic relationships of Trichostrongylus species in three counties of Kohgiloyeh and Boyerahmad (K-B) province, southwest Iran. Small intestine and abomasum of 70 sheep and goats were investigated. RESULTS: A total of 35 isolates of Trichostrongylus worms were detected and all were genetically identified as Trichostrongylus vitrinus. Analysis of 321 bp of the internal transcribed spacer 2 (ITS2) of ribosomal DNA revealed 16 genotypes. All genotypes were single nucleotide polymorphisms, including some hypervariable points. All sequences were trimmed to 170 bp, compared with sequences on GenBank including short sequences from other endemic foci of Iran and other countries and all isolates were used to generate a maximum likelihood phylogenetic tree, which consisted of two clades A and B. Clade A included isolates from Iran, Russia, New Zealand, Australia and the UK; clade B only contained South African isolates. Most clade A isolates (north, southwest and west Iran, Russia, New Zealand, Australia and UK) were in a similar phylogenetic position. One subclade was detected in clade A (isolates from Southwest Iran, New Zealand and UK). CONCLUSIONS: We hypothesize that drug resistant T. vitrinus may account for its exclusive detection in our samples. The high similarity of genotypes from Iran, New Zealand and UK may be due to their close political relationships during the colonial era. More research is needed to understand better the phylogeny of T. vitrinus and its relationship with drug resistance and human transmission.


Subject(s)
Phylogeny , Trichostrongylosis/veterinary , Trichostrongylus/classification , Animals , Feces/parasitology , Genotype , Goat Diseases/epidemiology , Goat Diseases/parasitology , Goats/parasitology , Humans , Intestine, Small/parasitology , Iran/epidemiology , Livestock , New Zealand , Sheep/parasitology , Sheep Diseases/epidemiology , Sheep Diseases/parasitology , Species Specificity , Trichostrongylosis/epidemiology , Trichostrongylus/isolation & purification , United Kingdom
4.
Parasit Vectors ; 13(1): 457, 2020 Sep 09.
Article in English | MEDLINE | ID: mdl-32907633

ABSTRACT

BACKGROUND: The worldwide increased difficulty to combat gastrointestinal nematode (GIN) infection in sheep, due to progressing anthelmintic resistance (AR), calls for an enhanced and standardized implementation of early detection of AR. This study provides a snapshot of the current AR status against benzimidazoles and macrocyclic lactones in southern Italy, generated with standardized techniques. METHODS: On 10 sheep farms, the efficacy of albendazole (ALB) and either eprinomectin (EPR) or ivermectin (IVM) was evaluated based on the faecal egg count reduction test (FECRT) performed with the Mini-FLOTAC. For each tested drug, 40 sheep were rectally sampled at D0 and sampled again 14 days after the treatment (D14). The FECRT was calculated from individual samples and pooled samples which consist of 5 individual samples. Efficacy was classified as 'reduced, 'suspected' and 'normal'. Coprocultures were set for D0 and D14 faecal samples of each group. From farms with FECR < 95%, an in vitro egg hatch test (EHT) and a follow-up FECRT using fenbendazole (FBZ) were conducted. RESULTS: Based on the FECR, high efficacy (from 95.7% to 100%) was observed for ALB and IVM in eight farms (Farms 3-10). On Farm 1 and Farm 2, the efficacy for the macrocyclic lactones was classified as 'normal', but 'reduced' efficacy was observed for ALB on Farm 1 (FECR = 75%) and 'suspected' efficacy on Farm 2 (FECR = 93.3%) with the predominant GIN genus Trichostrongylus followed by Haemonchus at D14. The FEC results of pooled samples strongly correlated with those of individual samples, for FEC at D0 (rs = 0.984; P < 0.0001) and at D14 (rs = 0.913; P < 0.0001). The classifications of efficacy in Farm 1 (FECR = 86.0%) and Farm 2 (FECR = 93.0%) in the follow-up FECRT with FBZ coincide with the main FECRT trial. The in vitro EHT confirmed AR in both farms (Farm 1: 89%; Farm 2: 74%). CONCLUSIONS: In regions like southern Italy, where the negative impacts from AR have played a minor role, efficient monitoring of AR is important in order to evaluate potential risks and being able to promptly respond with countermeasures.


Subject(s)
Anthelmintics/therapeutic use , Drug Resistance , Nematoda/isolation & purification , Nematode Infections/veterinary , Sheep/parasitology , Animals , Benzimidazoles/therapeutic use , Farms , Feces/parasitology , Fenbendazole/therapeutic use , Gastrointestinal Diseases/drug therapy , Gastrointestinal Diseases/parasitology , Gastrointestinal Diseases/veterinary , Haemonchus/drug effects , Haemonchus/isolation & purification , Italy , Ivermectin/analogs & derivatives , Ivermectin/therapeutic use , Nematoda/drug effects , Nematode Infections/drug therapy , Parasite Egg Count/methods , Sheep Diseases/drug therapy , Sheep Diseases/parasitology , Trichostrongylus/drug effects , Trichostrongylus/isolation & purification
5.
Acta Parasitol ; 65(3): 790-795, 2020 Sep.
Article in English | MEDLINE | ID: mdl-32319036

ABSTRACT

PURPOSE: The aim was to characterize the infection by Trichostrongylus spp. in patients from Chile using a combination of molecular detection techniques and phylogenetic analysis relating the findings to clinical and epidemiological reports of the patients METHODS: Strongylid eggs were detected in seven patients by coproparasitological techniques. From each sample a fragment of the ITS-2 ribosomal gene was amplified by PCR, sequenced and analyzed by the Neighbor-Joining method. RESULTS: All the sequences and phylogenetic clusters corresponded to T. colubriformis. Two samples presented a single nucleotide polymorphism showing two possible haplotypes. Six patients presented gastrointestinal symptoms. All of them lived on farms and used sheep manure as fertilizer. CONCLUSION: T. colubriformis was the strongylid involved in the infections of these Chilean patients associated with the presence of livestock and agricultural practices that favor infection by this type of nematode.


Subject(s)
Phylogeny , Rural Population , Trichostrongylosis/epidemiology , Trichostrongylus/genetics , Animals , Chile/epidemiology , DNA, Ribosomal Spacer/genetics , Feces/parasitology , Humans , Livestock , Parasite Egg Count , Polymorphism, Genetic , Sequence Analysis, DNA , Trichostrongylus/classification , Trichostrongylus/isolation & purification
6.
Mol Biochem Parasitol ; 236: 111260, 2020 03.
Article in English | MEDLINE | ID: mdl-31958470

ABSTRACT

Polymerase chain reaction followed by high resolution melting (PCR-HRM) analysis is a simple, rapid and accurate method for molecular detection of various nematode species. The objective of the present study was, for the first time, to develop a PCR-HRM assay for the detection of various animal Trichostrongylus spp. A pair of primers targeting the ITS-2 rDNA region of the Trichostrongylus spp. was designed for the development of the HRM assay. DNA samples were extracted from 30 adult worms of Trichostrongylus spp., the ITS-2-rDNA region was amplified using PCR, and the resultant products were sequenced and characterized. Afterwards, the PCR-HRM analysis was conducted to detect and discriminate Trichostrongylus spp. Molecular sequence analysis revealed that 24, 4, and 1 of the samples were T. colubriformis, T. vitrinus and T. capricola, respectively. Results from PCR-HRM indicated that complete agreement was relatively found between speciation by HRM analysis and DNA sequencing for the detection of Trichostrongylus species. The PCR-HRM analysis method developed in the present study is fast and low-cost; the method can be comparable with other molecular detection techniques, representing a reliable tool for the identification of various species within the Trichostrongylus genus.


Subject(s)
Polymerase Chain Reaction/methods , Trichostrongylus , Animals , DNA, Helminth/genetics , DNA, Ribosomal Spacer/genetics , Trichostrongylus/classification , Trichostrongylus/genetics , Trichostrongylus/isolation & purification
7.
BMC Infect Dis ; 20(1): 28, 2020 Jan 10.
Article in English | MEDLINE | ID: mdl-31924183

ABSTRACT

BACKGROUND: Parasitic trichostrongyloid nematodes have a worldwide distribution in ruminants and frequently have been reported from humans in Middle and Far East, particularly in rural communities with poor personal hygiene and close cohabitation with herbivorous animals. Different species of the genus Trichostrongylus are the most common trichostrongyloids in humans in endemic areas. Also, Ostertagia species are gastrointestinal nematodes that mainly infect cattle, sheep and goats and in rare occasion humans. The aim of the present study was to identify the trichostrongyloid nematodes obtained from a familial infection in Guilan province, northern Iran, using morphological and molecular criteria. METHODS: After anthelmintic treatment, all fecal materials of the patients were collected up to 48 h and male adult worms were isolated. Morphological identification of the adult worms was performed using valid nematode keys. Genomic DNA was extracted from one male worm of each species. PCR amplification of ITS2-rDNA region was carried out, and products were sequenced. Phylogenetic analysis of the nucleotide sequence data was performed using MEGA 6.0 software. RESULTS: Adult worms expelled from the patients were identified as T. colubriformis, T. vitrinus and Teladorsagia circumcincta based on morphological characteristics of the males. Phylogenetic analysis illustrated that each species obtained in current study was placed together with reference sequences submitted to GenBank database. CONCLUSIONS: The finding of current study confirms the zoonotic aspect of Trichostrongylus species and T. circumcincta in inhabitants of Guilan province. The occurrence of natural human infection by T. circumcincta is reported for the first time in Iran and the second time in the world.


Subject(s)
Trichostrongyloidea/genetics , Trichostrongyloidiasis/epidemiology , Trichostrongyloidiasis/transmission , Trichostrongylosis/epidemiology , Trichostrongylosis/transmission , Trichostrongylus/genetics , Zoonoses/epidemiology , Zoonoses/transmission , Animals , Anthelmintics/therapeutic use , Base Sequence/genetics , DNA, Helminth/genetics , DNA, Ribosomal/genetics , Feces/parasitology , Humans , Iran , Livestock/parasitology , Male , Phylogeny , Polymerase Chain Reaction , Prevalence , Trichostrongyloidea/isolation & purification , Trichostrongyloidiasis/drug therapy , Trichostrongylosis/drug therapy , Trichostrongylus/isolation & purification , Zoonoses/drug therapy
8.
Trop Anim Health Prod ; 52(4): 1699-1705, 2020 Jul.
Article in English | MEDLINE | ID: mdl-31865537

ABSTRACT

Co-infections caused by trypanosomes and gastro-intestinal nematodes (GINs) compromise cattle productivity and their control requires a holistic approach. The effectiveness of trypanocides and anthelmintics is compromised by increasing resistance. Use of combined chemotherapeutic products for synergy, mainly practiced in human medicine, is gaining importance in livestock. A trial to evaluate efficacy of VERYL®, containing diminazene diaceturate (3.5 mg/kg body weight) and levamisole chloride (5 mg/kg body weight) for the control of GINs in cattle, was conducted at KALRO-VSRI Muguga, Kenya, between June and August 2016. Thirty-eight cattle aged between 6 and 12 months, naturally infected with GINs, were randomly allocated into two groups; a treatment group received VERYL® intra-muscularly at 10 mL/100 kg bwt and a control group which received Veriben® (Diminazene aceturate) at 3.5 mg/kg bwt. Faecal egg counts (FECs), coproculture, packed cell volume (PCV) and local tolerance at the injection site were measured during the study. FECs were comparable between the treatment and control groups at day 0. However, treatment of cattle with VERYL significantly (p < 0.001) reduced FECs by day 7 and sustained to day 21 post-treatment. Coproculture results for the treatment and control groups revealed presence of Haemonchus, Cooperia, Ostertagia, Trichostrongylus and Oesophagostomum species. Cattle treated with VERYL® had a significant (p < 0.05) reduction in larval recoveries compared to the control group. VERYL® had minimal adverse effects which cleared after a short while and is thus recommended for controlling GINs in cattle.


Subject(s)
Anthelmintics/therapeutic use , Cattle Diseases/drug therapy , Diminazene/analogs & derivatives , Levamisole/therapeutic use , Nematode Infections/veterinary , Animals , Cattle , Diminazene/therapeutic use , Feces/parasitology , Haemonchus/isolation & purification , Kenya , Nematode Infections/drug therapy , Parasite Egg Count/veterinary , Random Allocation , Trichostrongylus/isolation & purification
9.
Parasitol Res ; 118(3): 891-900, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30719533

ABSTRACT

This study involved a national cross-sectional survey of gastrointestinal nematodes (GINs) of alpacas in Australia. A total of 1545 fresh faecal samples were collected from both sexes of alpacas and processed for faecal egg counts (FEC) and molecular identification of nematodes using the multiplexed tandem PCR assay. Based on egg morphology, the overall prevalence of GINs was 66% while that for strongyles was 59%. The overall mean FEC was 276 eggs per gram (EPG) of faeces, with the highest count of 17,415 EPG. Male alpacas had a higher prevalence (68%, 334/490) as well as mean FEC (328 ± 60 EPG) of GINs than females (63%, 602/954; 227 ± 26, respectively). Weaners had the highest prevalence (80%) whereas tuis had the highest FEC (402 EPG) of nematodes. The highest prevalence (77%, 293/383) and FEC (630 EPG) of GINs were observed in the summer rainfall zone followed by the Mediterranean-type rainfall, non-seasonal rainfall and winter rainfall zones. The characterisation of nematode DNA isolated from faeces revealed the occurrence of seven different GINs, including Camelostrongylus mentulatus, Cooperia spp., Haemonchus spp., Oesophagostomum spp., Ostertagia ostertagi, Teladorsagia circumcincta and Trichostrongylus spp. Besides, Nematodirus spp. and Trichuris spp. were also found during FECs. The prevalence of Haemonchus spp. was highest in the summer rainfall zone while that of C. mentulatus was highest in the Mediterranean-type rainfall, non-seasonal rainfall and winter rainfall zones. The findings of this study revealed that alpacas harbour many of the same nematodes as sheep and cattle.


Subject(s)
Camelids, New World/parasitology , Gastrointestinal Tract/parasitology , Nematode Infections/epidemiology , Nematode Infections/veterinary , Animals , Australia/epidemiology , Cattle , Cross-Sectional Studies , DNA, Protozoan/analysis , Feces/parasitology , Female , Haemonchus/isolation & purification , Male , Multiplex Polymerase Chain Reaction , Nematode Infections/parasitology , Ostertagia/isolation & purification , Parasite Egg Count , Prevalence , Seasons , Sheep , Sheep Diseases/parasitology , Surveys and Questionnaires , Trichostrongylus/isolation & purification
10.
Parasitol Res ; 118(3): 901-911, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30737673

ABSTRACT

We conducted a longitudinal survey on 13 alpaca farms in four climatic zones of Australia to understand the epidemiology of gastrointestinal nematodes (GINs) of alpacas. A total of 1688 fresh faecal samples were collected from both sexes of alpacas from May 2015 to April 2016 and processed for faecal egg counts (FEC) and molecular identification of eggs using the multiplexed-tandem PCR assay. Based on egg morphology, the overall prevalence of GINs was 61% while that for strongyles was 53%. The overall mean FEC was 168 eggs per gram (EPG) of faeces, with the highest count of 15,540 EPG. Weaners had the highest prevalence (73%) and mean FEC (295 EPG) of GINs followed by tuis, crias and adults. Alpacas in the winter rainfall zone had the highest prevalence (68%) as well as FEC (266 EPG) followed by Mediterranean-type, non-seasonal and summer rainfall zones. Trichostrongylus spp. (83%, 89/107), Haemonchus spp. (71%, 76/107) and Camelostrongylus mentulatus (63%, 67/107) were the three most common GINs of alpacas across all climatic zones. The mixed-effects zero-inflated negative binomial regression model used in this study showed that it could help to design parasite control interventions targeted at both the herd level and the individual alpaca level. The findings of this study showed that the epidemiology of GINs of alpacas is very similar to those of cattle and sheep, and careful attention should be paid when designing control strategies for domestic ruminants co-grazing with alpacas.


Subject(s)
Camelids, New World/parasitology , Gastrointestinal Tract/parasitology , Nematode Infections/epidemiology , Nematode Infections/veterinary , Animals , Australia/epidemiology , Cattle , Climate , Farms , Feces/parasitology , Female , Haemonchus/isolation & purification , Longitudinal Studies , Male , Multiplex Polymerase Chain Reaction , Nematode Infections/parasitology , Ostertagia/isolation & purification , Parasite Egg Count , Seasons , Sheep , Sheep Diseases/parasitology , Trichostrongylus/isolation & purification
11.
Vet Parasitol ; 261: 1-8, 2018 Sep 15.
Article in English | MEDLINE | ID: mdl-30253846

ABSTRACT

In this paper, we present for the first time a new tool, based on Droplet Digital™ Polymerase Chain Reaction (ddPCR), for absolute quantification of key genera of gastrointestinal (GI) nematode parasites of grazing livestock. Four combinations of primers/probe sets targeting the internal transcribed spacer region 2 (ITS2) of the ribosomal RNA gene array were designed using the Primer3 software, following in silico analysis of nucleotide sequences from nematodes of interest downloaded from common databases. The amplified regions include both a universal region for detection of any strongylid gastrointestinal parasite and three different genus specific regions, making it possible to differentiate between the most important GI nematodes of sheep in Sweden: Haemonchus, Teladorsagia and Trichostrongylus. Analysis of samples containing serial dilutions and different mixtures of genomic DNA extracted from different species of adult worms proved useful in assessment of different threshold settings with the QuantaSoft software. Analysis of template DNA from these worms indicated that ddPCR is a viable choice for detection and absolute quantification of the different genera and also in samples with multiple species. Interpretation of the ddPCR results was straightforward and choice of analytical approach had little influence on the final results. Thus, the results obtained in the different analytical approaches seemed to be robust and the concentrations determined were uniform. Furthermore, the linear range of the Haemonchus ddPCR assay was similar to that of real-time PCR (qPCR). Taken together, our data confirm the suitability of ddPCR for detection and absolute quantification of three major sheep pathogens when tested on larval cultures from pooled ovine faeces. The results also indicate that ddPCR can be a useful complement to applications based on conventional egg counting methods such as the faecal egg reduction test (FECRT).


Subject(s)
Haemonchiasis/veterinary , Ostertagiasis/veterinary , Polymerase Chain Reaction/veterinary , Sheep Diseases/diagnosis , Trichostrongylosis/veterinary , Animals , Feces/parasitology , Haemonchiasis/diagnosis , Haemonchiasis/parasitology , Haemonchus/isolation & purification , Ostertagia/isolation & purification , Ostertagiasis/diagnosis , Ostertagiasis/parasitology , Polymerase Chain Reaction/methods , Sheep , Sheep Diseases/parasitology , Sweden , Trichostrongylosis/diagnosis , Trichostrongylosis/parasitology , Trichostrongylus/isolation & purification
12.
Infect Genet Evol ; 63: 249-256, 2018 09.
Article in English | MEDLINE | ID: mdl-29705361

ABSTRACT

In this study, we characterize the diversity and estimated infection levels of gastrointestinal parasites circulating in two galago species, Galago demidoff and G. thomasi in two sites situated in the Southeastern forests of Gabon. Our study reveals that eleven parasites including nine helminthes (Ascaris spp., Ankylostoma spp., Dicrocoelium spp., Gongylonema spp., Oesophagostomum spp., Lemuricola spp., Strongyloides spp. Trichostrongylus spp. and Trichuris spp.) and two protozoans (Balantidium spp. and Entamoeba spp.) may infect Galago spp. with high infection rates. The results show that: a very similar parasite spectrum is found in both host species; all the taxa identified were previously observed in other Primate species and/or Man. They also show that age, gender and forest type may influence infection rates and/or parasite diversity found in a particular host and/or geographic area.


Subject(s)
Balantidiasis/veterinary , Entamoebiasis/veterinary , Galago/parasitology , Intestinal Diseases, Parasitic/veterinary , Nematode Infections/veterinary , Ancylostoma/classification , Ancylostoma/isolation & purification , Animals , Ascaris/classification , Ascaris/isolation & purification , Balantidiasis/epidemiology , Balantidiasis/parasitology , Balantidium/classification , Balantidium/isolation & purification , Dicrocoelium/classification , Dicrocoelium/isolation & purification , Entamoeba/classification , Entamoeba/isolation & purification , Entamoebiasis/epidemiology , Entamoebiasis/parasitology , Feces/parasitology , Female , Forests , Gabon/epidemiology , Intestinal Diseases, Parasitic/epidemiology , Intestinal Diseases, Parasitic/parasitology , Male , Nematode Infections/epidemiology , Nematode Infections/parasitology , Oesophagostomum/classification , Oesophagostomum/isolation & purification , Prevalence , Spiruroidea/classification , Spiruroidea/isolation & purification , Strongyloides/classification , Strongyloides/isolation & purification , Trichostrongylus/classification , Trichostrongylus/isolation & purification , Trichuris/classification , Trichuris/isolation & purification
13.
Am J Trop Med Hyg ; 98(3): 768-771, 2018 03.
Article in English | MEDLINE | ID: mdl-29313482

ABSTRACT

Sporadic cases of Tricostrongylosis are reported in humans. Diagnosis of enteric Trichostrongylus relies primarily on coproscopic analysis but morphological identification is difficult because of similarity among nematode species. The method is time consuming and requires some expertise. To overcome these limitations, we developed a molecular approach by real-time polymerase chain reaction (PCR) to provide a rapid, specific, and sensitive tool to detect Trichostrongylus spp. in human feces. We designed primers and probe specific for Trichostrongylus rDNA region 5.8S and internal transcribed spacer 2. Three Italian family clusters were analyzed and DNA sequencing was performed to confirm real-time PCR results comparing with known GenBank sequence data. Sequence analysis showed ≥ 99% identity to Trichostrongylus colubriformis and Trichostrongylus axei. This study provides a molecular methodology suitable for fast and specific detection of Trichostrongylus in fecal specimens and to distinguish the zoonotic species.


Subject(s)
Real-Time Polymerase Chain Reaction/methods , Trichostrongylosis/diagnosis , Trichostrongylosis/epidemiology , Trichostrongylus/genetics , Animals , Base Sequence , DNA Primers/chemical synthesis , DNA, Helminth/genetics , DNA, Helminth/isolation & purification , DNA, Ribosomal Spacer/chemical synthesis , Feces/parasitology , Humans , Italy/epidemiology , Sequence Alignment , Sequence Analysis, DNA , Sequence Homology, Nucleic Acid , Trichostrongylosis/parasitology , Trichostrongylus/classification , Trichostrongylus/isolation & purification
14.
Trop Anim Health Prod ; 50(4): 807-813, 2018 Apr.
Article in English | MEDLINE | ID: mdl-29280029

ABSTRACT

The objective of this study was to determine the prevalence of gastrointestinal nematode (GIN) infection in goat flocks on semi-arid rangelands of northeastern Mexico (25° N, 350-400 mm annual precipitation). The study included 668 pluriparous goats from 18 herds in five municipalities of Coahuila and Nuevo Leon, Mexico. Five genetic groups were considered (predominance of Boer, Nubian, Alpine, Saanen, and Toggenburg). Fecal samples were taken from the rectum of each animal to determine the number of eggs per gram (EPG) of GIN. The prevalence of flocks with GIN infections was 88.9%. Similar results were observed for the number of goats infected in the flocks. The Alpine breed presented the highest prevalence and highest EPG loads of GIN, whereas Boer and Nubian were the genetic groups with the lowest (P < 0.05) EPG. There was a negative effect of GIN infection on the live weight of goats (P < 0.05). The GIN genera found were Trichostrongylus spp. and Haemonchus spp. It was concluded that in the goat flocks of the semi-arid zones of Mexico was found a high prevalence of infections with gastrointestinal nematodes. The municipality and the breed of the animals were factors that showed influence on this prevalence and the level of infection of the goats.


Subject(s)
Gastrointestinal Diseases/veterinary , Goat Diseases/epidemiology , Trichostrongylosis/veterinary , Trichostrongylus/isolation & purification , Animals , Communicable Diseases , Feces , Female , Gastrointestinal Diseases/epidemiology , Goat Diseases/genetics , Goats , Haemonchus/isolation & purification , Mexico/epidemiology , Nematode Infections , Ovum , Parasite Egg Count/veterinary , Prevalence , Trichostrongylosis/epidemiology
15.
Parasitol Res ; 116(11): 3211-3219, 2017 Nov.
Article in English | MEDLINE | ID: mdl-29018987

ABSTRACT

This study evaluated patterns and species composition of parasitic infections detected over a 1-year period at an organic goat farm. As a result of coprological examination, the overall prevalence of observed strongylids (99%), coccidia of the genus Eimeria (98%), and Muellerius capillaris lungworms (93%) was calculated. The most prevalent strongylids recovered from incubated fecal samples were Haemonchus contortus (42%), genera Trichostrongylus (23%), Oesophagostomum columbianum (13%), and Teladorsagia circumcincta (11%). A maximum intensity of coccidia infection 5150 oocysts per gram, strongylids infection 9900 eggs per gram and lungworm infection 867.26 larvae per gram were detected. The various effects (including environment, host, and parasites) on milk yield, lactose, protein, and fat were evaluated using generalized linear mixed models. Milk yield (P < 0.0001), milk fat (P < 0.01), and lactose (P < 0.0001) were affected by month, i.e., these parameters were influenced by the month of the year, regardless of the individual goat. With the intensity of infection detected in our study, only protein content was affected (P < 0.01) by parasitic infection (exclusively caused by strongylids). Correlation between measurements from one individual revealed that the goat itself can substantially decrease protein content but has much less of an effect on fat, milk yield, and lactose. Based on our results, we can conclude that a low intensity of parasitic infections does not significantly affect milk yield and the qualitative parameters of milk.


Subject(s)
Goat Diseases/parasitology , Goats/parasitology , Haemonchus/isolation & purification , Metastrongyloidea/isolation & purification , Milk/metabolism , Oesophagostomum/isolation & purification , Trichostrongylus/isolation & purification , Animals , Czech Republic/epidemiology , Farms , Feces/parasitology , Female , Goat Diseases/epidemiology , Haemonchiasis/epidemiology , Haemonchiasis/veterinary , Lactation , Oesophagostomiasis/epidemiology , Oesophagostomiasis/veterinary , Organic Agriculture , Parasite Egg Count/veterinary , Seasons , Strongylida Infections/epidemiology , Strongylida Infections/veterinary , Trichostrongylosis/epidemiology , Trichostrongylosis/veterinary
16.
Acta Trop ; 176: 293-299, 2017 Dec.
Article in English | MEDLINE | ID: mdl-28760481

ABSTRACT

Human infections with Trichostrongylus species have been reported in most parts of Iran. The aim of this study was the identification, molecular characterization and phylogenetic analysis of human Trichostrongylus species based on ITS2 region of ribosomal DNA from Guilan Province, northern Iran. Stool samples were collected from rural inhabitants and examined by formalin-ether concentration and agar plate culture techniques. After anthelmintic treatment, male adult worms were collected from five infected cases. Genomic DNA was extracted from one male worm of each species in every treated individual and one filariform larva isolated from each case. PCR amplification of ITS2-rDNA region was performed and the products were sequenced. Among 1508 individuals, 46 (3.05%) were found infected with Trichostrongylus species using parasitological methods. Male worms of T. colubriformis, T. vitrinus and T. longispicularis were expelled from five patients after treatment. Out of 41 filariform larvae, 40 were T. colubriformis, and the other one was T. axei. Phylogenetic analysis showed that each species was placed together with reference sequences submitted to GenBank database. Intra-species similarity for all species obtained in the current study was 100%. T. colubriformis was found to be probably the most common species in this region of Iran. For the first time, the authors of the present study report the occurrence of natural human infection by T. longispicularis in the world. Therefore, the number of Trichostrongylus species infecting human in Iran now increased to ten.


Subject(s)
DNA, Helminth/genetics , DNA, Ribosomal/genetics , DNA, Ribosomal/isolation & purification , Feces/parasitology , Microfilariae/genetics , Phylogeny , Trichostrongylus/genetics , Animals , Base Sequence , Humans , Iran/epidemiology , Male , Microfilariae/isolation & purification , Polymerase Chain Reaction , Trichostrongylosis/epidemiology , Trichostrongylus/isolation & purification
17.
Acta Trop ; 173: 180-184, 2017 Sep.
Article in English | MEDLINE | ID: mdl-28595822

ABSTRACT

Trichostrongylus species remain one of the major health challenges in the tropical and summer rainfall regions worldwide. Identification of strongylid species diagnostic methods is vital for obtaining a deep understanding of the epidemiology, population biology, anthelmintic treatment efficacy, and drug resistance in order to design effective parasite control strategies. We evaluated a multiplex RE-PCR for the diagnosis of key Trichostrongylus spp. Genomic DNA amplification of Trichostrongylus colubriformis, Trichostrongylus axei and Trichostrongylus vitrinus was achieved as standard sample using specific primers located in the second internal transcribed spacer (ITSII) of nuclear ribosomal DNA (rDNA). The mentioned method was based on isolation of Trichostrongylus ova from human fecal samples using Willis method, the extraction of ova genomic DNA samples, followed by rDNA ITSII PCR and one-step multiplex RE-PCR using three restriction enzymes of HinfI, DraI, and MseI. The multiplex RE-PCR technique provides a useful tool for discriminating all Trichostrongylus spp., being useful for diagnostic, epidemiological, ecological studies, and control programs. This method is rapid, especially when numerous restriction enzymes are required for species differentiation or identification.


Subject(s)
Multiplex Polymerase Chain Reaction/methods , Trichostrongylosis/parasitology , Trichostrongylus/genetics , Trichostrongylus/isolation & purification , Animals , Anthelmintics , DNA Primers/genetics , DNA, Ribosomal/genetics , DNA, Ribosomal Spacer/genetics , Feces/parasitology , Humans , Time Factors , Trichostrongylosis/diagnosis
18.
Infection ; 45(2): 233-236, 2017 Apr.
Article in English | MEDLINE | ID: mdl-27796847

ABSTRACT

Trichostrongylus spp. are parasites that are seldom recognized as a cause of eosinophilia and gastroenteric symptoms in industrialized countries. The index of suspicion raises when several members of a same household present eosinophilia. We report four clusters of Trichostrongylus infection diagnosed in a single center, in northern Italy. Patients came from four different provinces of three Italian Regions. Some patients presented symptoms (abdominal pain and diarrhea were the most frequent ones, reported by 67 and 42% of our patients, respectively), while other were asymptomatic. All of them presented eosinophilia, that was severe (>5000 eosinophils/mmc) in 58% cases. Obtaining an accurate history from patients, investigating possible ingestion of vegetables contaminated by organic manure or sheep dejections, is particularly important to achieve diagnosis, also in light of the low sensitivity of parasitological tests.


Subject(s)
Cluster Analysis , Eosinophilia/epidemiology , Eosinophilia/etiology , Trichostrongylosis/epidemiology , Trichostrongylosis/pathology , Trichostrongylus/isolation & purification , Adolescent , Adult , Animals , Eosinophilia/diagnosis , Feeding Behavior , Female , Humans , Italy/epidemiology , Male , Trichostrongylosis/diagnosis , Young Adult
19.
Vet Parasitol ; 226: 88-92, 2016 Aug 15.
Article in English | MEDLINE | ID: mdl-27514891

ABSTRACT

Resistance to ivermectin and moxidectin was explored by a faecal egg count reduction test in two sheep flocks with suspected anthelmintic resistance. The FECRT confirmed one suspicion, with a mean percentage of reduction in egg excretion within the treated groups of 0% for ivermectin (CI 95%: -228 to 58) and 13% for moxidectin (CI 95%: -152 to 70). This was further explored by a controlled efficacy test. An experimental infection of 18 naïve lambs was set up using infective larvae isolated from this flock (5000 L3/lamb). Compared to the control group, abomasal worm burdens (Teladorsagia circumcincta) were reduced by 90% [CI 95%: 81.5-94.8] and 85% [CI 95%: 72.4-92.2] after ivermectin (p<0.05) and moxidectin (p<0.05) treatment respectively. Again, compared to the control group, there was a reduction for intestinal strongyles (Trichostrongylus colubriformis) of 100% and 99% [CI 95%: 97.5-99.7] for ivermectin and moxidectin respectively. No difference was found between the efficacy of moxidectin and ivermectin. Pharmacokinetic values indicated that the strongyles were submitted to anthelmintic concentrations usually lethal to them. This trial demonstrated the first multiple resistance of ovine strongyles in France.


Subject(s)
Antinematodal Agents/pharmacology , Ivermectin/pharmacology , Macrolides/pharmacology , Nematode Infections/veterinary , Sheep Diseases/drug therapy , Trichostrongyloidea/drug effects , Abomasum/parasitology , Animals , Antinematodal Agents/therapeutic use , Cecum/parasitology , Drug Resistance , Feces/parasitology , Female , France , Intestine, Small/parasitology , Ivermectin/therapeutic use , Macrolides/therapeutic use , Male , Nematode Infections/drug therapy , Nematode Infections/parasitology , Parasite Egg Count/veterinary , Sheep , Sheep Diseases/parasitology , Trichostrongyloidea/isolation & purification , Trichostrongyloidiasis/drug therapy , Trichostrongyloidiasis/parasitology , Trichostrongyloidiasis/veterinary , Trichostrongylus/drug effects , Trichostrongylus/isolation & purification
20.
J Wildl Dis ; 52(1): 126-30, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26745834

ABSTRACT

Coypus (Myocastor coypus) are widespread throughout Europe. In northern Italy, they are abundant in the flatland areas, and their high population densities can cause economic loss and ecosystem damage. We examined 153 coypus for selected parasitic and bacterial infections. We found Strongyloides myopotami (63.4% prevalence), Trichostrongylus duretteae (28.1%), Eimeria coypi (86.3%), and Eimeria seideli (6.8%), but did not find Giardia duodenalis or Cryptosporidium spp. We also isolated Staphylococcus aureus (10.1%), Escherichia coli (4.5%), and Streptococcus spp. (3.4%) from lung samples; no Salmonella spp. were isolated from fecal samples. Coypus had antibodies to Toxoplasma gondii (28.9%) and to four serovars of Leptospira interrogans (44.9%); Australis/Bratislava was the serovar most frequently detected. It is clear that coypu can be infected with pathogens of human and veterinary importance.


Subject(s)
Bacterial Infections/veterinary , Parasitic Diseases, Animal/epidemiology , Rodent Diseases/epidemiology , Rodentia/parasitology , Animals , Bacterial Infections/epidemiology , Bacterial Infections/microbiology , Coccidiosis/parasitology , Coccidiosis/veterinary , Eimeria/isolation & purification , Escherichia coli/isolation & purification , Escherichia coli Infections/epidemiology , Escherichia coli Infections/microbiology , Escherichia coli Infections/veterinary , Feces/microbiology , Feces/parasitology , Italy/epidemiology , Lung/microbiology , Parasitic Diseases, Animal/parasitology , Prevalence , Risk Factors , Rodent Diseases/microbiology , Rodent Diseases/parasitology , Rodentia/microbiology , Staphylococcal Infections/epidemiology , Staphylococcal Infections/microbiology , Staphylococcal Infections/veterinary , Staphylococcus aureus/isolation & purification , Streptococcal Infections/epidemiology , Streptococcal Infections/microbiology , Streptococcal Infections/veterinary , Streptococcus/isolation & purification , Strongyloides/isolation & purification , Strongyloidiasis/epidemiology , Strongyloidiasis/parasitology , Strongyloidiasis/veterinary , Trichostrongylosis/epidemiology , Trichostrongylosis/parasitology , Trichostrongylosis/veterinary , Trichostrongylus/isolation & purification , Urban Population
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