<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.     

Pathological,serological, and virological findings in sheep infected simultaneously with <Emphasis Type="Italic">Bluetongue</Emphasis>, <Emphasis Type="Italic">Peste-des-petits-ruminants</Emphasis>, and <Emphasis Type="Italic">Sheeppox viruses</Emphasis>

In this study, pathological, serological and virological examinations were performed on 15 sheep from a flock of 250 sheep and lambs that suffer from simultaneous naturally occurring BTV, PPRV and SPV outbreaks. SPV was diagnosed macroscopically and histopathologically, BTV was diagnosed by ELISA, and PPRV was diagnosed pathologically and by ELISA. Clinically fever, diarrhea, depression, polypnea, conjunctivitis, lacrimation, rhinitis, erosive stomatitis, edema of eyelids, photophobia, cutaneous eruption with erythematous areas especially noticeable in wool-free parts of the body and axilla lesions evolving into papules were observed. At necropsy, the most effected organs were lungs and gut. Subepicardial hemorrhages were also commonly seen. While typical pox lesions were observed in some lambs, usually fibrinous pleuropneumonia was more prominent lung lesion. SPV and PPRV lesions were seen at the histopathological examination of the lesioned tissues, BT lesions were mild than SPV and PPRV microscopically. Serum and leukocyte samples of 15 animals were examined for PPRV and BTV by ELISA; 5 samples were positive for PPRV and 6 BTV, 4 were positive for both PPRV and BTV simultaneously. One hundred animals died, most were lambs. Mortality rates were 100% in lambs and 80% in the herd.     

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.     

Prevalence of thin-walled <Emphasis Type="Italic">Sarcocystis cruzi</Emphasis> and thick-walled <Emphasis Type="Italic">Sarcocystis hirsuta</Emphasis> or <Emphasis Type="Italic">Sarcocystis hominis</Emphasis> from cattle in Iran

Bovine sarcocystosis is caused by Sarcocystis cruzi and is known to cause considerable morbidity and mortality in cattle. This species is distributed worldwide in cattle and is the most prevalent of the Sarcocystis species infecting cattle. There is high infection rate of sarcocyst in cattle in Iran, but to our knowledge, there is no study about identification of Sarcocystis species. This work aimed to survey prevalence of S. cruzi cyst in slaughtered cattle of Isfahan, Iran. In this study, esophageal and diaphragmatic muscles of 100 cattle were collected from Fesaran abattoir of Isfahan and examined for the presence of Sarcocystis spp. cysts macroscopically and microscopically. No macroscopic sarcocysts were found in any of the samples. In light microscopy, 89 out of 100 cattle (89%) had thin-walled cysts of S. cruzi, while 21 out of them (21%) had thick-walled sarcocysts. In addition to light microscopy, ultrastructural features of the thin-walled cyst confirmed the presence of S. cruzi.     

<Emphasis Type="Italic">Mycobacterium bovis</Emphasis>, but also <Emphasis Type="Italic">M. africanum</Emphasis> present in raw milk of pastoral cattle in north-central Nigeria

Using deletion typing technique, five mycobacteria isolated from unpasteurised milk samples from cows in north-central Nigeria were characterized as Mycobacterium bovis (n = 4) and M. africanum (n = 1). This report emphasizes that transmission between the animal and human reservoir is a serious threat in Nigeria.     

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.     

Immunoprotection of recombinant <Emphasis Type="Italic">Eg</Emphasis>.P29 against <Emphasis Type="Italic">Echinococcus granulosus</Emphasis> in sheep

Objective

This study aims to investigate the immunoprotection of recombinant Eg.P29 (rEg.P29) vaccine and analyze the underlying mechanism in sheep.

Methods

Three groups of male sheep were immunized subcutaneously with rEg.P29 and PBS, Freund’s complete adjuvant as controls, respectively. After prime-boost vaccination, the sheep were challenged with encapsulated Echinococcus granulosus eggs. The percentage of protection in sheep was determined 36 weeks after the infection. Humoral immune response was analyzed for specific IgG, IgG1, IgG2, IgM and IgE levels. Moreover, cytokines including interferon (IFN)-γ, interleukin (IL)-2, IL-4,and IL-10 were also evaluated.

Results

Immunization with rEg.P29 induced protective immune responses up to 94.5 %, compared with immunoadjuvant group. The levels of specific IgG, IgG1, IgG2, and IgE as well as IFN-γ, IL-2, and IL-4 significantly increased after two immunizations (P < 0.05); however, the levels of IgM and IL-10 did not show difference.

Conclusion

rEg.P29 showed Immunoprotection and induced Th1 and Th2 immune responses; hence, rEg.P29 is a potential vaccine for E. granulosus infection.

     

Control of <Emphasis Type="Italic">Haemonchus contortus</Emphasis> in sheep using basidiocarps of <Emphasis Type="Italic">Agaricus blazei</Emphasis> Murril

Objective

This study evaluated the effects in vitro and in vivo of Agaricus blazei against Haemonchus contortus in sheep.

Methods

The in vitro efficacy of aqueous extract on egg hatching inhibition (EHI) was investigated and after 72 h incubation with varying concentrations the effects on, blastomeres, embryonated eggs, and first stage larvae (L1) were evaluated. Larval development inhibition (LDI) for dry powder and the aqueous extract were evaluated in fecal cultures of sheep infected with H. contortus. In vivo efficacy was determined by reduction in fecal egg count (FEC). Lambs were treated with powder A. blazei (11.4 g/kg pc) or trichlorfon, or were untreated and the possible toxicity of this fungus was monitored by plasmatic enzyme analysis.

Results

Concentrations equal to and higher than 3.62 mg/mL and of aqueous extract were 100% effective in the EHI test. In the LDI test, LC90 was estimated for 5.66 and 106.0 mg/g fecal culture for aqueous extract and powder, respectively. The mean FEC in lambs 14 days post-treatment with A. blazei powder was significantly lower than observed for the negative control, and the serum levels of aspartate transaminase and alanine transaminase were normal.

Conclusion

The fungi supplementation promotes, respectively, high and moderate anthelmintic efficacy with in vitro and in vivo tests, respectively, suggesting it as an alternative or complementary treatment for haemonchosis in sheep.

     

Experimental infection of <Emphasis Type="Italic">Peste des Petit Ruminant</Emphasis> virus and <Emphasis Type="Italic">Mannheimia haemolytica</Emphasis> A2 in goats: immunolocalisation of <Emphasis Type="Italic">Mannheimia haemolytica</Emphasis> antigens

Nigerian strain of Peste des Petit Ruminant (PPR) virus and Mannheimia haemolytica (MH) biotype A serotype 2, was used successfully to reproduce a concurrent disease in West African Dwarf goats. The development of the various pathological features were studied at regular intervals following infection. The acute inflammatory reaction which had developed by day 3 after initial infection was characterised by flooding of the alveoli by neutrophils, oedema, hemorrhage and syncytial cells together with a moderate bronchial and bronchiolar epithelial necrosis. This progressed to a milder acute broncho interstitial pneumonia with giant cells. At this stage, the mucosal immunity were well developed especially the aggregate form of NALT and more of nodular forms of BALT. The organisms were demonstrated with strong immunostaining in the necrotic center, necrotic alveolar wall, fibrin, serous exudate, and degenerated leukocyte in the alveoli and respiratory airways. The bacterial antigens were observed as a strong immunostaining in the blood vessels of the nasal septum, sinusoid in the liver and interstium of the kidney, cytoplasm of alveolar macrophages, pneumocytes, bronchial and bronchiolar epithelium, in the monocytes in the blood vessels. These findings confirmed the enhancement of MH tropism especially in the respiratory tract, liver and kidney. It also showed that West african dwarf goats are highly susceptible to the intratracheal combined infection of PPR virus and MH. The fact that the infection induces strong mucosal responses, this phenomenon can be explored in Africa with the use of combined PPR virus and MH intranasal vaccines to curtail the menace of pneumonia associated with the combined infection on field.     

Seroprevalence of <Emphasis Type="Italic">Brucella</Emphasis> antibodies in camels in Katsina State,Nigeria

A cross-sectional study was carried out to determine the status of Brucella infection in one-humped (Dromedary) camels in the North and Central senatorial districts of Katsina State, Nigeria. Nine hundred and eighty serum samples from live and slaughtered camels were tested. Modified Rose Bengal plate test (RBPT) and serum agglutination test (SAT) with ethylenediaminetetraacetic acid, (EDTA) were used as screening and standard tests, respectively. The prevalence of Brucella antibodies were 110 (11.2%) and 103 (10.5%) for RBPT and SAT, respectively. Of the 472 and 508 serum samples tested from the herds and abattoirs, respectively, 63 (13.3%) and 47 (9.3%) were positive by RBPT while 62 (13.1%) and 41 (8.1%) were positive by SAT, respectively. Based on the results, it was concluded that Brucella antibodies were present in camels in the study area. Poor management practices and mixing of camels with other species of livestock as well as unrestricted movement of camels were proposed to be the reasons for the prevalence of the disease in the study area. In view of the public health importance of the disease, it is recommended that there is the need to develop a strategic plan to decrease spread of brucellosis in the study area.     

Anticoccidial effects of <Emphasis Type="Italic">Aloe secundiflora</Emphasis> leaf extract against <Emphasis Type="Italic">Eimeria tenella</Emphasis> in broiler chicken

Anticoccidial effects of Aloe secundiflora crude leaf extract was tested in broiler chickens following oral infection with Eimeria tenella. Sixty 22-day-old birds were divided into six groups of ten birds each. Three treatment groups A, B, and C were fed with the extract (100, 250, and 500 mg/day, respectively) mixed in feed for 10 days, and three control groups: group D (drug control) administered 300 mg/l of sulfachloropyrazine sodium soluble powder in drinking water for 5 days, group E (infected/non-medicated positive control), and group F (uninfected/non-medicated negative control). Except for group F, all groups were orally inoculated with 75,000 sporulated oocysts of E. tenella. The effects of the extract on E. tenella infection were evaluated by severity of bloody diarrhea, body weight (BW) gain, oocyst output, and lesion score. No bird in the treated groups died of coccidiosis, and severity of bloody diarrhea was milder than in the positive control group. BW gains in the treated groups were significantly higher than in group E (p < 0.05). The lesion scores of the treated groups were significantly lower than that of group E. Oocyst output in groups A, B, and C were 11.23, 8.24, and 6.82 × 10⁶, respectively. As compared with the negative control group (12.84 × 10⁶), the reductions in oocyst production were 12.54, 35.83, and 46.88%, respectively. Oocyst output significantly reduced with an increase in Aloe dosage. The findings of this study suggest that Aloe secundiflora extract presents an alternative anticoccidial agent for the control of avian coccidiosis.     

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.     

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.     

Investigation of seroprevalence of <Emphasis Type="Italic">Theileria equi</Emphasis> and <Emphasis Type="Italic">Babesia caballi</Emphasis> in horses in Nigde province,Turkey

The prevalence of equine piroplasmosis caused by Theileria equi and Babesia caballi in Nigde, in central Anatolia, Turkey has remained unknown. Serum samples were obtained from a total of 125 horses and were tested for antibodies to T. equi and B. caballi using the Indirect Fluorescence Antibody Test (IFAT). Twenty-three (18.4%) horses were seropositive for equine piroplasmosis. Anti-T. equi was observed in 16 horses (12.8%) while anti-B. caballi was detected in 12 horses (9.6%). In addition, 5 serum samples were positive for both parasites. The prevalence rates of antibodies to T. equi and B. caballi for female and male horses were statistically indifferent (p = 0.19 and 0.90). The difference between the seropositivity rates to T. equi among age groups was statistically insignificant (p = 0.44) while the difference to B. caballi among age groups is statistically significant (p = 0.01). Seropositivity rates ranged from 2.9% to 25.7% for T. equi and 2.9% to 14.3% for B. caballi from the selected districts in Nigde. A statistically significant difference on seropositivity rates for the study sites was observed for only T.equi (p = 0.03). This study indicates that T. equi is higher than B. caballi in Nigde. This study was supported by the Scientific Research Projects Unit of Nigde University (FEB 2007/08).