Survey of <Emphasis Type="Italic">Sarcocystis</Emphasis> infection in slaughtered cattle in Kerman,Iran

The parasite of genus Sarcocystis is one of the most commonly found parasite in domestic animals worldwide. Some species of Sarcocystis cause important economic loss when causing clinical and sub clinical disease. The aim of this study was to determine the prevalence of Sarcocystis in slaughtered Cattle in Kerman, Iran. The prevalence of Sarcocystis spp. infection was investigated in 480 cattle, slaughtered from May 2005 to February 2006 in the Kerman, Iran using naked eye examination for macroscopic Sarcocysts, and peptic digestion, muscle squash, squeezing methods for microscopic types. Muscles from heart, tongue, and esophagus, cervical and abdominal muscles of 480 slaughtered cattle were examined for Sarcocystis cysts. The prevalence of microscopic Sarcocystis cysts in cattle was detected in 100% and there was no macroscopic cyst in examined cattle.     

Prevalence and molecular analysis of Sarcocystis infections in cattle in Northwest Iran and the first global report of S. gigantea in cattle

Cattle are intermediate host for several species of Sarcocystis, including S. cruzi, S. hirsuta, and S. hominis with high prevalence worldwide. The present study aimed to determine the prevalence of Sarcocystis infection, species identification, and phylogenetic analysis of the parasite in cattle in Northwest Iran. The samples of diaphragm and esophagus from 290 cattle were collected from slaughterhouses in Northwest Iran and subjected to macroscopic, microscopic, and histopathology examinations, PCR-RFLP, sequencing and phylogenetic analyses. Tissue cysts of Sarcocystis spp. were detected in 92% of cattle by digestion and microscopic tests. Based on the PCR–RFLP and specific PCR, 87.9% and 1.03% of isolates were identified as S. cruzi, and S. hominis, respectively. Macrocyst was seen in a single sample that was identified as S. gigantea. The haplotype network exhibited the extension of the various haplotypes of S. cruzi between neighboring provinces in Northwest Iran. Heterogeneity analysis of S. cruzi 18S-rRNA sequences indicated genetic diversity among S. cruzi isolates (Haplotype diversity: 0.733-0.854) consisting 16 haplotypes; however, the nucleotide differences showed low diversity (0.01481 to 0.03351). Pair wise sequence distance matrix amongst S. cruzi sequences indicated an intra-species divergence of 0%-7.8% and identity of 92.6%-100%. Sarcocystis infection is highly prevalent in cattle in Northwest Iran, with the predominance of S. cruzi, and genetic variants of this species are unequivocally distributing in Northwest provinces. First global detection of S. gigantea in cattle reflects new insights of transmission dynamic and biology of this parasite in Iran.     

Prevalence and distribution patterns of <Emphasis Type="Italic">Sarcocystis</Emphasis> spp. in buffaloes in Beni-Suef,Egypt

This study was performed for the purpose of investigating the prevalence of Sarcocystis spp. in buffaloes in Beni-Suef Governorate, Egypt. Both macroscopic (Sarcocystis fusiformis) and microscopic (Sarcocystis levinei) cysts were recognized, and were differentiated by their morphological features and location in the tissues. Of 379 buffaloes examined in abattoirs in Beni-Suef, 299 were found to be infected, with an overall prevalence of 78.9%. Depending on age, three categorized groups of naturally infected buffaloes were examined: male buffalo calves aged 1.5–2 years, adult females aged 2–5 years, and females older than 5 years. Among these groups, infection rates were 74.5%, 82.3%, and 81.2%, respectively. Organs examined included esophagus, tongue, and heart. Macroscopic cysts were examined by the naked eye through meat inspection in abattoirs, while the pepsin-digestion method and the histological technique were applied to detect microscopic cysts. It has been found that esophagus showed the highest rate of infection among the infected organs, with both macroscopic and microscopic cysts seen in the infected buffaloes. Moreover, results of the pepsin-digestion method proved more accurate than those produced by the histological technique in terms of infection rates for the microscopic cysts. Our findings indicated that infected buffaloes aged 2–5 years showed the highest mixed infection rate (82.3%) for both types of cysts. The high prevalence of microscopic Sarcocystis spp. in Beni-Suef Governorate reflects a significant role played by stray dogs, rather than cats, in the transmission of these parasites.     

Detection of <Emphasis Type="Italic">Mycobacterium avium</Emphasis> subsp. <Emphasis Type="Italic">paratuberculosis</Emphasis> in tissue samples of cattle and buffaloes

Tissue samples were collected at random from cattle (Bos taurus) and buffalo (Bubalus bubalis) from an abattoir of the district of Lahore and were analyzed for the presence of Mycobacterium avium subsp. paratuberculosis and Mycobacterium bovis through acid-fast staining and polymerase chain reaction (PCR). Body condition of animals and diarrhea were recorded. Most of the animals were emaciated. Diarrhea was noticed in 15.6% of buffaloes and 19.2% of cattle. Intestinal pathology was observed in 29% of buffaloes and 32.8% of cattle. Number of mesenteric lymph node (MLN) showing gross lesions was a bit higher (35.6%) in cattle than buffalo (31.2%). Acid-fast staining of tissue scraping smears revealed the presence of acid-fast bacilli (AFB) in 17.4% intestinal and 16.4% MLN tissue samples in buffalo, while in cattle 19.2% intestinal and 17.8% MLN were found positive for AFB. In buffaloes, PCR confirmed 12.8% intestinal and 12.4% MLN positive samples for M. avium subsp. paratuberculosis. However, in cattle, PCR analysis demonstrated 14.2% positive results for M. avium subsp. paratuberculosis in both MLN and intestinal tissue samples. PCR also confirmed M. bovis in 5.8% of cattle and 5% of buffalo MLN and intestinal tissues. PCR positive tissue samples for M. avium subsp. paratuberculosis were from those animals which were emaciated, having diarrhea, and severe gross lesions. AFB were also detected in tissue scraping smears of these animals. It is concluded that infection by various mycobacterium species can be differentiated by PCR, which is not possible by acid-fast staining technique.     

A new PCR-RFLP method for detection of <Emphasis Type="Italic">Anaplasma marginale</Emphasis> based on 16S rRNA

Species of the genus Anaplasma (Rickettsiales: Anaplasmataceae) are obligate intracellular tick borne pathogens. Three species of Anaplasma that infect cattle and sheep (A. marginale, A. centrale and A. ovis) are well recognized. Of these erythrocytic Anaplasma, A. marginale can cause diseases in the livestock with high economical losses. Species-specific PCR based on 16S rRNA gene is commonly used for detection of Anaplasma species but can not differentiate A. marginale, A. centrale and A. ovis because of sequence similarity. In this study DNA extraction was performed on 50 blood samples with presence of Anaplasma spp. in marginal point of erythrocytes in their blood smears. The extracted DNA from blood cells was analyzed by PCR and PCR-RFLP using primers derived from 16S rRNA gene and restriction endonuclease Bst1107 I. The restriction endonuclease Bst1107I only recognizes the sequence (GTATAC) in corresponding PCR product of A. marginale and cut it. The nucleotide sequence of the A. marginale 16S rRNA gene was determined and compared with the sequences of A. marginale in GenBank. The 16S rRNA of A. marginale in Iran was completely similar to the related sequence deposited in GenBank at accession number of M60313. In the present study we propose a new PCR-restriction fragment length polymorphism analysis (RFLP) method based on 16S rRNA gene for specific detection of A. marginale.     

Occurrence of anti-<Emphasis Type="Italic">Toxoplasma gondii</Emphasis> antibodies in female cattle in south-west of Iran

Toxoplasmosis, caused by Toxoplasma gondii, is a significant disease in livestock and humans. In Iran, studies shows that T. gondii infection in humans is relatively high and prevalence is associated mainly with consumption of undercooked meat or meat products. We have examined 450 serum samples from female cattle distributed over all Ahvaz, the center of Khouzestan province, south-west of Iran. IgG antibodies to T. gondii were assayed by the modified agglutination test using whole tachyzoites of T. gondii, and found in 71 (15.77%) of 450 cattle with titers of 1:25 in 38, 1:50 in 18, 1:100 in 11, 1:200 in three and 1:400 in one. Titers of antibodies were decreased in cattle over 2 years old. These results indicate that T. gondii infection in cattle of Khouzestan is relatively considerable, but not very high and consumption of beef may be a source of infection for humans in south-west of Iran.     

<Emphasis Type="Italic">Theileria</Emphasis> (<Emphasis Type="Italic">Babesia</Emphasis>) <Emphasis Type="Italic">equi</Emphasis> and <Emphasis Type="Italic">Babesia caballi</Emphasis> Infections in Horses in Galicia,Spain

The control of equine piroplasmosis is becoming increasingly important to maintain the international market open to the horse industry. The purpose of this study was to demonstrate the occurrence of equine piroplasmosis (Theileria equi and Babesia caballi) in Galicia, north-west Spain, and to compare haematological and serum biochemistry parameters between non-parasitaemic horses and horses parasitaemic with T. equi and B. caballi. Sixty serum samples (control group) were taken from healthy horses pastured on two farms, and examined for evidence of equine T. equi and B. caballi infection by indirect fluorescent antibody test (IFAT). Of the 60 samples, 24 (40%) and 17 (28.3%) samples were positive for T. equi and B. caballi, respectively. Twelve (20%) samples were positive for both parasites. Haematology and serum biochemistry were compared between controls and a series of 36 horses clinically affected by T. equi (25) or B. caballi (11). Compared with the healthy group, there was a 43% and 37% decrease in the haematocrit for T. equi and B. caballi infection, respectively. Parasitaemic horses presented an intense anaemia and serum biochemistry signs of liver damage. The anaemia was more severe in T. equi-infected than in B. caballi-infected horses. Our results suggest that equine piroplasmosis is widespread in the region and is a cause for concern.     

Antimicrobial resistant <Emphasis Type="Italic">Salmonella</Emphasis> in dairy cattle in the United States

Increased frequency of antimicrobial resistant Salmonella isolated from humans over the last quarter century in the United States has led to concern about the contribution animal production systems have played in the emergence and spread of antimicrobial resistant Salmonella. In order to better understand the potential role of dairy cattle as a reservoir for antimicrobial resistant Salmonella, it is important to understand methods currently used to measure the prevalence of antimicrobial resistance among Salmonella from human and animal populations. This review describes the biology of Salmonella and antimicrobial resistance, methods used to monitor antimicrobial resistance, and studies that have measured the prevalence of antimicrobial resistant Salmonella among human and dairy cattle populations in the U.S. Although the prevalence of antimicrobial resistance among Salmonella from healthy dairy cattle is low, similar trends in the prevalence of resistance among Salmonella from clinically ill human and dairy cattle populations were observed in the literature.     

Seroprevalence of <Emphasis Type="Italic">Toxoplasma gondii</Emphasis> infection in yaks (<Emphasis Type="Italic">Bos grunniens</Emphasis>) in northwestern China

The seroprevalence of Toxoplasma gondii infection in yaks (Bos grunniens) was surveyed in Qinghai Province, northwestern China in May and June 2010. A total of 650 serum samples were collected from six counties and assayed for T. gondii antibodies by an indirect hemagglutination test. Antibodies to T. gondii were found in 35.08% (228/650) with the highest rate of 55.34% in Chengduo County. The results of the present survey indicated that infection with T. gondii in cattle is widely spread in China, including yaks in Qinghai Province.     

Virulence of <Emphasis Type="Italic">Brucella abortus</Emphasis> isolated from cattle and water buffalo

Brucellosis has been documented in domestic water buffalo (Bubalus bubalis) but published literature is limited despite the importance of this species in tropical agricultural systems. The objective of this study was to compare the virulence of Brucella abortus isolates recovered from cattle and water buffalo. Nineteen strains of B. abortus from cattle and domestic water buffalo in Trinidad were intraperitoneally inoculated into BALB/c mice. Spleens were cultured for B. abortus and histopathological severity scores were calculated based on lymphoid depletion, lymphoid necrosis, splenitis, and macrophage accumulation. A general linear model approach was used to estimate the effect of isolate source (cattle versus water buffalo) on virulence. Isolates of water buffalo origin were significantly less virulent in the mouse model based on recovered B. abortus from splenic tissues, spleen/weight ratio, and lymphoid necrosis but not overall histopathological severity scores. Further investigation of isolates recovered from water buffalo might provide the key to the development of procedures for brucellosis control in tropical environments.     

<Emphasis Type="Italic">Streptococcus uberis</Emphasis>-specific T cells are present in mammary gland secretions of cows and can be activated to kill <Emphasis Type="Italic">S. uberis</Emphasis>

The presence, phenotype and function of Streptococcus uberis-specific T cells in the mammary gland secretion (MGS) and blood of cows exposed to S. uberis were assessed. MGS T cells in the udder were purified and incubated with autologous blood monocytes as antigen-presenting cells (APC). Most cows, irrespective of prior S. uberis infection status and lactation status, were shown to have S. uberis-specific T cells both in MGS and in the blood. When cells from a subgroup of cows were studied, it was found that the S. uberis-specific T cells produced high levels of interferon-gamma (IFN-γ), but low levels of interleukin-10 (IL-10). A high percentage of responding T cells were of the CD8 ⁺ memory (CD45RO) subset. T cells from the MGS specific for S. uberis were propagated from animals during the drying off period and expanded in vitro using interleukin-2 (IL-2) and S. uberis antigens. This led to the accumulation of T cells of the CD8 ⁺ subset bearing memory cell markers (CD45A ⁻ , CD45RO ⁺ ), which released high levels of IFN-γ. Four of the five T cell lines derived from the MGS of three animals had substantial direct killing activity towards S. uberis in vitro. It is concluded that there is an emergence of S. uberis-specific bactericidal T cells in the MGS of cows after infection or environmental exposure to S. uberis. Vaccines aimed at activating and expanding this T cell population in the mammary glands of cattle may offer an avenue for the prevention of mastitis caused by S. uberis.     

The analysis of <Emphasis Type="Italic">groEL</Emphasis> gene in <Emphasis Type="Italic">Salmonella enterica</Emphasis> subspecies <Emphasis Type="Italic">enterica</Emphasis> serovar Typhimurium isolated from avians by PCR-Restriction Fragment Length Polymorphism method

Salmonella enterica subspecies enterica serovar Typhimurium causes food-borne outbreaks and systemic diseases in humans and animals. groEL gene (also known as mopA gene in S. Typhimurium), possessing conserved sequence, plays an important role in invasion of bacteria. The purpose of present study was to identify the polymorphism of groEL gene among different avians in different regions by PCR-RFLP method. Fifty two S. Typhimurium isolates (Broiler (n = 13), Layer (n = 12), Duck (n = 5), Goose (n = 5), Sparrow (n = 8), Canary (n = 3), Pigeon (n = 5) and Casco parrot (n = 1). were identified using serotyping as well as multiplex-PCR. Then, amplification of groEL gene performed and amplified products subjected to restriction digestion with BsuRI enzyme. Three RFLP profiles, A, B and C, generated DNA fragments between approximately 100–1,000 bp in size, were observed. The RFLP profile A was observed in 35 (67.3%), profile B in 14 (26.9%) and profile C in 3 (5.77%) of isolates. S. Typhimurium isolates recovered from 13 broilers (two of which profile A, 9 profile B and 2 profile C) and from 8 sparrows (two of which profile A, 5 profile B and 1 profile C) showed all three profiles, but 12 layers and other avians (including Canary (n = 3), Goose (n = 5), Duck (n = 5), Pigeon (n = 5) and Casco parrot (n = 1)) showed profile A. None of these profiles was allotted for a special region. The result of present study showed that S. Typhimurium undergoes genetic mutations in groEL gene under unpleasant milieu in different regions and in different avians. Thus, genetic diversity, despite conserved nature of groEL gene in S. Typhimurium, may exist but it depends on the condition where bacteria have settled. To our knowledge, three RFLP profiles of groEL gene generated by BsuRI restriction enzyme were not reported previously.     

Biological control of <Emphasis Type="Italic">Ascaris suum</Emphasis> eggs by <Emphasis Type="Italic">Pochonia chlamydosporia</Emphasis> fungus

Ascaris suum is a gastrointestinal nematode parasite of swines. The aim of this study was to observe Pochonia chlamydosporia fungus on biological control of A. suum eggs after fungus passage through swines gastrointestinal tract. Eighteen pigs, previously dewormed, were randomly divided into three groups: group 1, treated with the fungus isolate VC4; group 2, treated with the fungus isolate VC1 and group 3 did not receive fungus (control). In the treated groups, each animal received a 9 g single dose of mycelium mass containing P. chlamydosporia (VC1 or VC4). Thereafter, animal fecal samples were collected at the following intervals: 8, 12, 24, 36, 48, 72 and 96 h after treatment beginning and these were poured in Petri dishes containing 2% water-agar culture medium. Then, 1,000 A. suum eggs were poured into each dish and kept in an incubator at 26°C and in the dark for 30 days. After this period, approximately 100 eggs were removed from each Petri dish and morphologically analyzed under light microscopy following the ovicidal activity parameters. The higher percentage observed for isolated VC4 eggs destruction was 57.5% (36 h) after fungus administration and for isolate VC1 this percentage was 45.8% (24 h and 72 h) (p > 0.01). P. chlamydosporia remained viable after passing through the gastrointestinal tract of swines, maintaining its ability of destroying A. suum eggs.     

Ovicidal activity of seven <Emphasis Type="Italic">Pochonia chlamydosporia</Emphasis> fungal isolates on <Emphasis Type="Italic">Ascaris suum</Emphasis> eggs

The ovicidal effect of the nematophagous fungus Pochonia chlamydosporia on eggs of Ascaris suum was tested under laboratory conditions. A. suum eggs were plated on 2% water–agar with seven fungal isolates (Isol. 5, Isol. 31, Isol. 1, VC1, Isol. 12, Isol. 22 and VC4) and control without fungus. After 5, 7, 10, 14, 15 and 21 days of incubation, approximately 100 eggs were removed from the plates and classified according to the following parameters: type 1, biochemical and physiological effect without morphological damage to the eggshell, type 2, lytic effect with morphological alteration of the eggshell and embryo and type 3, lytic effect with morphological alteration of eggshell and embryo showing hyphal penetration and internal egg colonization. The isolates effectively destroyed A. suum eggs and all types of effects were observed during the experiment. There was no variation in ovicidal capacity (type 3 effect) among the isolates (p > 0.05) throughout the experiment. After 21 days, isolate 5 showed the highest percentages of type 3 effect (58.33%). The results indicated that P. chlamydosporia (Isol. 5, Isol. 31, Isol. 1, VC1, Isol. 12, Isol. 22 and VC4) can destroy A. suum eggs and is, therefore, a potential biological control agent of nematodes.     

Identification and characterization of a <Emphasis Type="Italic">Streptomyces</Emphasis> sp. isolate exhibiting activity against multidrug-resistant coagulase-negative Staphylococci

The resistance of 220 coagulase-negative Staphylococci (CNS) (associated with animal disease) to 13 antibiotics were determined using the disk diffusion method. 35.9% of multidrug-resistant coagulase-negative Staphylococci (MR-CNS) exhibited resistance to five or more than five antibiotics; all of these bacteria were resistant to methicillin too. The new Streptomyces sp. ABRIINW111 was isolated from the Zagros Mountains Hamadan, Iran. The 16S rDNA sequence of the isolate indicated that it has 98% similarity to S. levis, but some mutations in the alpha and gamma regions of the 16S rDNA sequence emphasize the probability of the existence of a new species. Preliminary and secondary antibacterial screenings revealed that the isolate is active against gram negative and positive bacteria. The diethyl ether extracted metabolite of the Streptomyces sp. ABRIINW111 showed an effective antibacterial activity against MR-CNS. So the diethyl ether extract of the new Streptomyces sp. strain ABRIINW111 can inhibit the MR-CNS in vitro, and it can offer a new approach to treat MR-CNS infectious patients.     

Re-emergence of multi-drug resistant <Emphasis Type="Italic">Salmonella enterica</Emphasis> serovar Stanley from cattle

During 2009, Salmonella enterica subspecies enterica serovar Stanley isolates were recovered from cattle diagnostic specimens in southern Japan, and the isolates were examined to characterize the genetic determinants involved in this new pathogenicity that associated with mortality in cattle. All the isolates were multi-drug resistance exhibited resistance to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, oxytetracycline, and kanamycin (ACSSuT-Km) encoded by bla _TEM, catA, aadA1, sul1, tet(A), and aphA1 genes, respectively. Class 1 integrons of 1.5-kb size were detected in all MDR isolates. The isolates harboured easily transferable plasmids of ca. 210-kb with the potential of transmitting resistance phenotype and genotype detected in the donor isolates. XbaI-digested PFGE patterns generated two related clusters implicated in the dissemination of multi-drug resistance amongst Salmonella Stanley isolates. An emergence of multi-drug resistant Salmonella Stanley amongst food-producing animals, including cattle is a threat to human health, as resistant isolates may be transmitted to humans through the food chain.     

Host Switch of <Emphasis Type="Italic">Lamellodiscus elegans</Emphasis> (Monogenea: Monopisthocotylea) and <Emphasis Type="Italic">Sparicotyle chrysophrii</Emphasis> (Monogenea: Polyopisthocotylea) between Cage-reared Sparids

Sharpsnout bream (Diplodus puntazzo) has been used in Adriatic aquaculture for less than a decade, but the decreasing trend of rearing this species will probably result in its complete substitution by more exploited sea bream (Sparus aurata). Only two facilities still rear both fish species in neighbouring cages in monoculture. A switch of parasites was observed between sparids during monitoring of the gill monogeneans of farmed fish. In wild fish of the Adriatic Sea, Lamellodiscus elegans (Monogenea: Monopisthocotylea) has previously been reported in annular (Diplodus annularis), and two-banded sea bream (D. vulgaris) and sharpsnout bream (D. puntazzo), and the present study confirmed its presence also in sea bream, in low prevalence and abundance. The exclusively sea bream monogenean Sparicotyle chrysophrii (Monogenea: Polyopisthocotylea) was also isolated from sharpsnout bream, showing prevalence and abundance values even higher than in its resident host. In the occurrence of L. elegans in sea bream, the opportunistic switch resulted in lower abundance and prevalence than in the original host, while in the second case of switching the monogenean S. chrysophrii showed better reproductive capacity on a new host (sharpsnout bream). Both cases point to the possible enlargement of parasite host range.     

Occurrences of thermophilic <Emphasis Type="Italic">Campylobacter</Emphasis> in cattle slaughtered at Morogoro municipal abattoir,Tanzania

An investigation was conducted in Morogoro municipality to assess the likelihood of slaughter cattle posing public health risk of contaminating carcasses with thermophilic Campylobacter. Butchers and meat shopkeepers were interviewed on source of slaughter cattle, method of animal and carcass transportation, carcass dressing, meat storage facilities, access to clean water and availability of food hygiene practices. Faecal samples were collected from 107 slaughter cattle and after slaughter; four different parts of dressed carcasses (i.e. from ham, neck, pelvis and thigh muscles) were also sampled. In addition 107 cattle meat samples for Campylobacter culture were collected in different meat shops. The samples were subjected to standard bacteriological examination using Skirrows protocol. It was found that cattle slaughter, dressing and meat handling in meatshops was done under unhygienic condition. Thermophilic Campylobacter prevalence in slaughter cattle was 5.6% while contamination rate of dressed carcasses and cattle meat at shops was 9.3% and 1.9%, respectively. The majority of thermophilic Campylobacter isolated were C. jejuni (88.9%) while C. coli was isolated at 11.1%. Findings of this study suggest possibility of humans acquiring zoonotic Campylobacter infections from cattle meat particularly when meat preparation and processing is not done properly. More work is required to establish the magnitude of zoonotic enteric Campylobacteriosis in humans and epidemiological role of cattle and other animals in the study area.     

Sero-prevalence and identification of <Emphasis Type="Italic">Ornithobacterium rhinotracheale</Emphasis> in broiler flocks in south-eastern Iran

Ornithobacterium rhinotracheale is a gram negative bacterial pathogen causing respiratory tract infections in poultry. Tracheal, lung and serum samples were obtained from 21 broiler flocks of 8 farms from a slaughterhouse located in south-eastern of Iran. Among 630 tracheal and lung samples from samples resulting from 315 chickens, 11 (3.5%) ORT isolates were identified using biochemical tests. The isolates originated from 9 (42.9%) flocks out of 4 farms. All of the isolates were recovered from tracheal swabs and showed an API 20NE identification biocode 0-2-2-0-0-0-4. Of the 420 serum samples examined by ELISA, 134 (31.9%) sera from 17 (81.0%) flocks were positive for ORT antibodies. These results indicate that ORT is present in most broiler flocks with respiratory disorders in southeast Iran.