sedigheh nabian

The Ichthyophthirius multifiliis (Ich) is one of the most important parasites in aquaculture industry, which causes the fatal disease of Ichthyophthiriasis. Treatments for this parasite include the use of certain chemical medications. However, due to the many negative effects of chemical compounds on the environment and humans, extensive studies of plant extract effectiveness in treating this parasite are very important. This study evaluated the alcoholic pomegranate (Punica granatum) peel extract's antiparasitic activity against the parasite I. multifiliis in vitro. Under laboratory conditions, the anti-parasitic efficacy of pomegranate peel alcoholic extract against I. multifiliis at doses of (0.5, 1, 2, 4, and 8 g/l) was evaluated after exposure for six hours. Additionally, this extract's toxicity was assessed for 96 hours on zebrafish at dosages of 0.5, 1, 2 and 4 g/liter. The collected data were statistically contrasted with the results of the positive control sample (15 ppm formalin) and the negative control treatment. The study revealed that the theronts may be destroyed in 6 hours by concentrations of 2, 4, and 8 g/liter, in addition to the concentration of 1 g/liter, which caused the death of 96% of theronts within this period. In the toxicity test, the concentrations (2, 4, and 8 g/L) were highly toxic, and all the fish died, but the concentrations of 1 and 0.5 g/L were safe doses within 96 hours. Also, the value of EC50 in this research was calculated as 1.41 g/liter. Therefore, the alcoholic extract of pomegranate peel in doses of 2, 4, and 8 g/l is not effective for clinical treatment and is only suitable as an antiseptic. So, the dose of 1 g/l has very good results and is recommended for clinical treatment.

The mites of the genus Tropilaelaps are parasites of honey bee brood. Their Feeding on haemolynph of bee larvae and pupae causes brood malformation, death of bees, and subsequent colony decline or absconding. Their development takes about 1 week and the mites are scattered on the body of the bees. There are at least four species in the genus Tropilaelaps, Each species tends to be associated with a particular bee species. Two species (T. clareae and T. mercedesae) are damaging pests of Apis mellifera. The other two species (T. koenigerum and T. thaii) appear to be harmless to A. mellifera.In this article, an attempt has been made to carefully review the Tropilaelaps mite as a newly emerging parasite from various aspects including the characteristics of the parasite, its life cycle, distribution in the world, methods of diagnosis, treatment, and its potential in the country. It is hoped that by paying special attention to this pest, the entry and spread of this parasite in the country will be prevented and a step forward will be taken toward to progress of beekeeping in the country.

Leishmania is a vector-borne protozoon, which causes visceral, cutaneous and mucocutaneous leishmaniosis in human and animals. Monocyte- derived exosome vaccines can be used as prophylaxis and immunotherapy strategies. The aim of this study was to design a multiple-epitope candidate vaccine using leishmaniolysin (GP63) and rK39 proteins against Leishmania major and L. infantum for monocyte-derived exosome preparation.

This study was carried out in Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran, 2023–2024. Effective immunodominant epitopes were selected from two antigenic proteins of GP63 and rK39 using various immunoinformatics and bioinformatics approaches. Vibrio cholerae β- subunit was used as an adjuvant to stimulate immune responses. Then, appropriate linkers were selected for the fusion of epitopes. The 3D model of candidate vaccine was predicted and validated.

This designed candidate vaccine could effectively be used as a prophylaxis strategy against leishmaniosis.

A candidate vaccine was designed using bioinformatic and immunoinformatic studies with virtual acceptable quality; however, effectiveness of this vaccine should be verified through further in-vitro and in-vivo studies.

BACKGROUND:

 Honey bee venom contains complex compounds such as polypeptides, enzymes, and amines. One of the important components of bee venom is the phospholipase A2 enzyme, which is considered an important honey bee venom allergen and is also used to treat some diseases. This enzyme is found in other insects, arachnids, snakes, and mammalian cells, and its function is the hydrolysis of the second ester bond of glycerophospholipids and the release of fatty acids and lysophospholipids. Although transient transfection can produce recombinant proteins, stable cells are more suitable for high-scale production with economic efficiency.

The present study created a stable cell line to produce recombinant phospholipase A2 from honey bee (Apis mellifera) venom.

Plasmid cloning DNA vector containing phospholipase A2 gene was prepared by Macrogen Company. The recombinant plasmid was transferred to Chinese hamster ovary cells by heat shock method, and gene expression was carried out in a HamsF12 culture medium containing neomycin antibiotic. After increasing polyclonal strains containing plasmid, monoclonal clones were selected by limiting dilution. Then, monoclonal clones were propagated, the soup of the selected cells was collected and concentrated, and the protein expression was checked by sodium dodecyl-sulfate polyacrylamide gel electrophoresis test.

The results of electrophoresis, which was performed to confirm the expression of the phospholipase A2 gene in the cell soup, showed a band with a molecular weight of 20 kilodaltons, which confirms the creation of a stable cell line for the production of recombinant phospholipase A2 honey bee venom.

After the transient transfection of the plasmid containing this gene, several cells undergo recombination due to having repair mechanisms and putting the desired gene along with the antibiotic resistance gene in their genome. These cells can be selected and propagated by adding antibiotics to the culture medium.

Enzymatic digestion of extra cellular matrix proteins by proteinases of Leishmania promastigotes is a complex process. Hence, studies on functional proteomics of these enzymes can help select these enzymes as possible vaccine candidates or selecting candidates for chemotherapy and immunotherapy. Several proteolytic enzymes are involved in virulence of Leishmania spp. These enzymes are mostly serine, cysteine and metalloproteases. We aimed to detect proteases in Leishmania promastigote exosomes.

Serine, cysteine and metalloproteases were investigated in exosomes and lysate of L. major promastigote using gelatin zymography. The study was carried out in the Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran, in 2021.

Zymography findings of metalloproteinases showed transparent bands, including a 63-kDa glycoprotein (GP63). This glycoprotein is a major surface metalloproteinase. In addition, transparent bands belonged to serin proteases and cathepsin were demonstrated in gels associated to Leishmania promastigote lysate and exosomes.

Several metalloproteases, serin proteases and cathepsins were shown in promastigote lysate and exosomes of L. major, which could purified and used as fractions for immunodiagnostic.

Management of severe acute respiratory syndrome coronavirus 2 in humans depends on the availability of vaccines or effective drugs. Studies have shown that angiotensin‑converting enzyme 2 (ACE2) is responsible for binding the viral spike glycoproteins to human cells. Melittin from the bee venom of Apis melifera is a peptide with antimicrobial activities.

In this study, important amino acid residues of ACE2 interacting with spike glycoproteins of the virus were described based on the ACE2‑spike–glycoprotein interface. This has been previously analyzed by Robson in crystal structures of the receptors and ligands. Flexible linkers and 31 amino acid residues from N‑terminal of ACE2 as coronavirus spike binding domains (SBDs) were added to 17 N‑terminal amino acids of melittin (the hydrophobic motif) to construct a hybrid peptide or M‑ACE2SBD. Then, secondary and tertiary structures of the peptide were predicted.

Docking of the hybrid peptide and coronavirus SBDs was carried out as well. Previous studies showed that toxicity and hemolytic activity of the melittin hydrophobic motif decreased in comparison to native melittin due to the lack of peptide binding to the exposed anionic lipids of the human cell membranes and hence the novel peptide can be recommended as an appropriate drug for clinical uses.

This study has hypothesized that 17 N‑terminal amino acids of the mutant melittin used in M‑ACE2SBD design are potentially hydrophobic and attached coronavirus‑2 through lipid envelope of the virus.

Leishmaniasis is an infectious and parasitic disease that is among the most important zoonotic diseases in tropical and subtropical regions, as well as developing countries. Despite the efforts of clinical researchers over the years, few signs of progress have been made in the treatment of cutaneous leishmaniasis leading to satisfactory clinical improvement. In this study, activated melittin was used to treat leishmaniasis caused by Leishmania major. Moreover, liposomes and nanoparticles of albumin were used separately for the entry of myelin into leishmania-infected macrophages. Furthermore, their effects on Leishmania-infected cells were compared in vivo.

After designing activated melittin peptide for a selective expression using pep fold and expasy servers, liposomes and albumin nanoparticles were used to transfer the peptide into the cell and evaluate the function of transfer vectors on Leishmania major-infected cells. Finally, they were investigated in vivo to analyze the anti-leishmaniasis activity of carriers containing activated melittin and their uptake by macrophages.

After wounding the BALB/c mice on the second week, an effective concentration of 100 μl of each of the nanoparticles of albumin and liposome containing 25 μg of activated melittin was injected into BALB/c mice as a therapeutic dose. After the fifth week, in the third group (control), the wound size increased significantly and eventually led to the death of the mice in the control group. However, in the first and second groups treated with liposomes containing peptide and nanoparticle albumin, at the beginning of the fifth week, the wound size got smaller, and the recovery process initiated (P<0.05).

The results of this part of the study showed that the therapeutic effects with liposome carriers and albumin nanoparticles containing 25 μg/ml activated melittin in mice with leishmaniasis can be compared with those of the control group. Liposome and nanoparticle carriers of albumin-containing activated melittin have been suggested as an effective, appropriate, and even alternative treatment for cutaneous leishmaniasis.

Rhipicephalus (Boophilus) spp. are important vectors for Babesia and Anaplasma species causing severe economic losses in livestock. Chemical compounds are commonly used to control tick infestation; however, acaricides resistance in tick has led to move toward alternative strategies such as vaccination. In this study, we introduced a vaccine candidate, namely CaTro against Rh. microplus tick composing of immunogenic B-cell epitopes derived from Rh. microplus cathepsin L and tropomyosin proteins. To evaluate this vaccine candidate, firstly the CaTro sequence was inserted into the prokaryotic expression vector and the recombinant protein CaTro was expressed in Bl21 bacteria. Afterward, purification was performed by Ni-NTA affinity chromatography. The quality of purified recombinant CaTro was also analyzed using sodium dodecyl sulfate-gel electrophoresis and western blotting. Moreover, to evaluate the induction of immune response, the rabbits were immunized with purified recombinant protein combined with Freund’s adjuvant. The findings of this study revealed molecular weight of expressed protein (CaTro) as 38.00 kDa. Furthermore, anti-CaTro antibody was detected in immunized rabbit's sera through dot blotting; while, there was not any response to the control rabbit's sera. The results suggest that CaTro is a potential candidate to develop an anti- Rh. microplus tick.

Leishmaniasis is characterized by strong inflammatory responses with high levels of inflammatory cytokines that induce microRNA 21 and matrix metalloproteinases. Melittin has inhibitory effects on proliferation of various cells via induction of apoptosis. Melittin can be integrated in cell membranes and induce apoptosis. Thus, designation of biomolecules for the selective destroy of the infected cells is a treatment option. One approach is the precise engineering of constructs for the selective expression of melittin in the infected cells.

For this aim we designed a construct composing melittin nucleotide sequence and nucleotide sequence coding for polyanionic peptide function inhibitory element to further guarantee the selective function of melittin in inflamed tissues and infected cells, were included in a construct as melittin inhibitor via matrix metalloproteinase degradable linker.

Reverse complementary sequences were designed so melittin sequences for the selective targeting of Leishmania could be expressed in infected cells using cell microRNA machinery.

Translation machinery in infected cells with increased miR-21 could translate melittin, MMP linker and polyanionic inhibitor through a non-canonical pathway. Then, the MMP linker is degraded and selective killing of Leishmania infected cells would happen.

The vastness of the Ferula assa-foetida L. habitats in Kerman province is significantly high in comparison to other provinces. For the local people therefor, it is important to improve its production and marketing as a natural resource economy source. In this study randomly 19 experienced experts in the field of medicinal plants of Ferula assa-foetida L. were questioned. After conducting face-to-face interviews and completing the questionnaire, data on the production and marketing of Ferula assa-foetida L. gathered in the strengths. They mostly asked to focus on the, weaknesses, strengths, opportunities and threats. Answers were put in SOWT matrix and ranked, to get the production and marketing strategies. ANP method also was used. Results showed that the most important strength point is the optimal quality of the Ferula assa-foetida L. resin, the most important weakness is the lack of government support, the most important opportunity is the marketing aboard and the most important threat is the smuggling and destruction of the rangeland habitats of this plant. On the basis of the result conversion of the raw material in to industrial is suggested.

Crimean–Congo hemorrhagic fever (CCHF) is a fatal disease caused by Nairovirus classified within the Bunyaviridae family. The virus is transmitted to humans through the bites of infected ticks or direct contact with viremic animals or humans. The current study aimed to detect the virus genome in ticks from Khorasan Razavi Province.

One hundred hard ticks were collected randomly from 100 sheep in four different areas of the province. Collected ticks were kept alive and identified. All the ticks were analyzed for the presence of CCHF virus genome using reverse transcriptase polymerase chain reactions (RT-PCR).

The identified ticks were belonging to Hyalomma marginatum (16% female and 6% male), Rhipicephalus turanicus (52% female and 25% male), and Dermacentor raskemensis (1%). The CCHF virus genome was found in Hyalomma marginatum (5% male from Taibad and Sabzevar region and 1% female from Taibad). Genetic analysis of the virus genome isolated from two regions (Sabzevar and Taibad) showed 100% identity.

This study indicated that CCHF should be regarded as a risk-borne infection in this province. Therefore, special health management is needed to control this disease.

Bee venom contains various biomolecules, such as enzymes, peptides, and amines. The immune sys-tem produces IgG antibodies against bee venom proteins. However, IgE antibodies may also be developed in allergic individuals.

In this study, immune responses, including IgG and IgE reactions to bee venom were assessed in vari-ous individuals, using the immunoblotting technique.

Serum samples were collected from 20 people of three major groups, namely beekeepers, allergic individu-als, and normal people. Venom samples of honey bees and wild bees were collected from the suburbs of Tehran, Iran. Furthermore, commercial honey bee venom samples extracted from Apis mellifera and samples of wild bees extracted from Polistes and Vespula were purchased from France. Immunoblotting was carried out using the sera of subjects and anti-human IgG and IgE coupled to horseradish peroxidase.

The results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed similar protein bands in Iranian and European honey bee venoms, including α-glucosidase (170 kDa), Api m (100 kDa), acid phosphatase (49 kDa), hyaluronidase (43 kDa), phospholipase A2 (17 kDa), and melittin (2 kDa). In wild bees, two bands were found with the molecular weights of 35 and 25 kDa belonging to antigen 5 and phospholipase A1, respectively. These were not observed in honey bee venoms. Immunoblot analysis revealed that all the mentioned proteins were immunogenic and al-lergenic in different individuals. Hyaluronidase, as well as phospholipases A1 and A2, were the major allergens in most individuals, while IgE reaction to melittin was only reported in one person.

In conclusion, studies on antibodies against bee venoms can be useful in immunotherapy. Different people indicated distinct allergenic patterns. Therefore, further similar assays are recommended before, during, and after immunotherapy.

Ticks are one of the most important ectoparasites in animals that cause economic losses in livestock industry. So, removal or reduction of ticks on animals is necessary. Cysteine proteases are among the compounds that play an important role in the physiological action of ticks and are a good candidate for the anti-tick vaccine. Cathepsins is one of the most important cysteine proteases.

The aim of this study was cloning and expression of recombinant cathepsin L gene of Rhipicephalus annulatus in order to evaluate its immunogenicity.

After collection the ticks were cultivated. Then RNA was extracted from ticks, cDNA was synthesized by using specific primer of cathepsin and amplification by RT-PCR. The desired genes were cloned into expressional pQE30 plasmid. Further, a shorter sequence of the cathepsin gene (654 bp) was prepared as a synthetic plasmid. The expression of the protein produced by both recombinant plasmids in the E.coliBL21 prokaryotic expression system is carried out and the immunity of the recombinant proteins was evaluated by Dot Blot and Western Blot using serum of challenged rabbits with recombinant protein and calves infected with ticks were examined and compared.

The results of this study indicate that the protein derived from recombinant plasmid No. 2 had higher expression and purity due to its solubility. Also, the challenge of rabbit serum with these proteins was able to identify both recombinant proteins. But the serum of challenged calves with ticks did not show a satisfactory response with recombinant proteins.

Although the sera reaction of calves infested with ticks was lower than the challenged rabbits sera with cathepsin L, this result was expected, because L cathepsin protein is considered as a concealed antigen.  Overall, the recombinant cathepsin L could be an appropriate candidate for immunizing calves against Rhipicephalus annulatus, although it seems further investigations are necessary.

Colony Collapse Disorder is a mysterious phenomenon in which worker bees abruptly disappear from a beehive. The aim of this study was to estimate the prevalence of CCD and its relation to climate and Nosema spp infections. This Cross sectional study was done from April to September 2016. With respect to different climatic zones of the country, a total of 183 ‎apiaries were selected. In each apiary, 5 percent of the colonies were ‎randomly sampled. Adult bee samples were then examined for the presence of Nosema infections using PCR. Data were analyzed by Chi-square using SPSS version 21.0. The results showed the prevalence of colony collapse disorder in the studied apiaries at 26.8%. The CCD prevalence was ‎20.5% in ‎humid, ‎16.1% in semi humid‎, ‎22.7% in very humid, 38.2% in arid, ‎43.8% in semi-‎arid and ‎16% in Mediterranean conditions. Comparing CCD phenomenon in different climatic regions, there were significant differences (P<0.05). The prevalence of Nosema ceranae infection was 85 (46.4%), however, infection with Nosema apis was not observed in the samples either in pure form or as ‎associated infection. There was no statistical significant difference between symptomatic and asymptomatic apiaries with colony collapse disorder in terms of presence or absence of N.ceranea (P>0.05).   The results suggest that climate could influence the prevalence of Colony Collapse Disorder. It may be due to different foraging resources in under studied area. According to findings of this study it seems that N.ceranea alone cannot be the cause for CCD. Further studies are needed to clarify the interactions between climate and other possible causes.

Argas persicus has a great importance for health and veterinary, it can transmit many infectious agents such as Borrelia anserina (avian spirochetosis) and Aegyptianella pullorum. Distinguishing Argasidae due to close morphological relationship is difficult. In the present study, we performed molecular analyses based on PCR and sequencing of Amplicon derived from 16S rRNA and COX1 genes of A. persicus specimens in several provinces of Iran.     Out of seventy Argas persicus collected and confirmed morphologically, eight ticks were chosen from five provinces of Iran for gene analysis. Their DNA were extracted and amplificated using primers derived from 16 S ribosomal RNA and COX1 genes using PCR. Then the amplicons were sequenced and analyzed by Chromas software and sequence alignment program (Clustal W). Phylogenetic analysis was also conducted using MEGA ver. 6.06 with a maximum-likelihood method. Sequencing results indicated that all eight samples belonged to A. persicus species. Their nucleotide sequencing revealed that the interspecific sequence differences of both genes (16S rRNA genes and COX1) between our isolates were very infrequent. All isolates from different provinces were conserved across regions except for one isolate that exhibited a difference of only 1 nucleotide. Within Phylogenetic tree, A. persicus formed a clade with A. persicus from other regions of the world (South Arica, Italy, China, and South Australia). Our findings suggested a very close phylogenetic relationship between A. persicus specimens obtained from different regions of Iran. Keywords:

Rhipicephalus (Boophilus) annulatus is an important tick, which can transmit parasites and bacteria to cattle. It can also cause hides damage, growth reduction and sometimes paralysis. So, the tick infestation can be regarded as a major problem in the livestock industry. Different control methods including the chemical ones are used to fight ticks, but because of developing resistance to chemical treatments, researchers try to find some immunogenic proteins for vaccine production. Investigating these tick proteins could be an important step in the identification of biological molecules for the purpose of developing control strategies. The aims of present study were to evaluate and analyze Boophilus annulatus larval extract proteins by 2- dimensional gel electrophoresis patterns (proteomic profiling) and identification of some its immunogenic proteins by two dimensional immunoblotting and MALDI TOF/TOF mass spectrometry referred to as Mazandaran strains. The steps followed here were: tick preparation and culture, 1-D electrophoresis, and then 2-D electrophoresis, Western blotting and Mass spectrometry and then Mass data were analyzed by Mascot software. Analysis of the produced 2D image identified approximately 80 protein spots with different Molecular weight and PI by Coomassi blue staining. Based on immunogenicity (through Western blotting) and high concentration, 10 protein spots (between 14 and 97 kDa) candidates for MALDI TOF and MALDI TOF - TOF MS. Among the 10 proteins spots, Vitellogenin, Vitellogenin-2 precursor, tropomyosin, hypotethical protein ISCW001652 were identified proteins with immunogenic properties. Also, quantification analysis showed some proteins had more quantity in soluble larvae protein extract and some such as vitellogenin had some isoforms. The results of this study could be a preliminary step towards selecting proteins candidated to develop vaccines against ticks.

Coccidiosis causes morphologic alteration in intestinal mucosa resulting in reduction of absorptive surface. Anticoccidials used as feed additives may induce changes in the intestinal mucosa. This study was designed to assess intestinal morphometry in broilers infected with Eimeria under different anticoccidial treatments. To evaluate the effect of salinomycin and amprolium+ethopabate on intestinal morphometry in broilers experimental coccidiosis, in Tehran, Iran in May 2015, fifty-four Ross 308 birds were randomly divided into two challenged and unchallenged groups at the age of 12 days. The birds were challenged with Eimeria field isolate at day 14. Different growth and parasitological parameters including weight gain, feed consumption, FCR, macroscopic lesion score and oocyst score were recorded 7 d post-inoculation. Histological sections from four main parts of intestine (anterior, middle, lower intestines and cecum) were prepared and analyzed. Villus width and length and total mucosal thickness were measured microscopically. Amprolium+ethopabate and salinomycin significantly reduced coccidiosis gross lesions in infected birds. Microscopically anticoccidial administration in the presence of infection has significantly increased the villus length while the presence of amprolium+ethopabate in the absence of infection has greatly increased the mucosal thickness and villi height in comparison to the control group. Anticoccidials may induce some histological changes in the mucosa when there is no parasite to be affected. Some of these effects may be advantageous for the intestinal epithelium integrity and hence the birds’ performance.

Blastocystis is the most common anaerobic protozoa living in the large intestine of a broad spectrum of vertebrates.
The aim of this study was to investigate the Blastocystis infection rate in inflammatory bowel disease (IBD) patients.
A total of 80 stool samples were collected from IBD-proved patients. All stool samples were cultivated in Dulbecco’s modified Eagle’s medium and checked by light microscopy for detection of Blastocystis. The Correlation between demographic data of IBD patients and Blastocystis was calculated using SPSS 23.
The enrolled patients comprised of 52 (65%) men and 28 (35%) women. The study showed Blastocystis in 16/80 (20%) of the samples by microscopic examination and culture method. The parasite was seen among 12 (23.08%) and 4 (14.29%) men and women, respectively. No statistically significant correlation was found between infection with the parasite and animal contact. Fisher’s exact test represented that there was no correlation between gender and the presence of Blastocystis (p value= 0.397). Fisher’s exact test denoted that there was no statistical correlation between age and the presence of the parasite (p value= 0.130).
In this study, Blastocystis was found in 20% of enrolled patients who suffered from IBD. This infection rate was significantly higher than the studies have previously described Blastocystis in this group of patients.

Regarding emerging tick resistance against acaricides, researches have been shifted toward alternative approaches such as immunologic methods. Vaccine preparation is an alternative way in which choosing appropriate protein with high immune induction potency is a prerequisite. In addition according to studies, using more than one protein could better enhance the immune induction and antibody production. Choosing immunogenic epitopes from selected proteins and adjoining them with a suitable linker is one of the novel approaches in vaccine design.

Based on the fact that both cathepsin and tropomyosin proteins of Rhipicephalus tick were previously recognized as potent immunogenic antigens, we predicted the immunogenic epitopes of these proteins by immunoinformatic methods. Among studied epitopes, those that were met by multiple bioinformatics tools were used.

Finally, the polytopic construction was designed by assembling the selected epitopes and connecting them with linkers.

Using immunoinformatic tools, we predicted the characteristics of two genes of Rhipicephalus annulatus tick larva as fused potent vaccine candidates namely, cathepsin and tropomyosin.

Many wild-caught reptiles harbor some kind of parasites. Captivity with negative effect of poor sanitary and husbandry management may lead to clinical disease. The increasing trend in keeping non-native reptile species in the last decade emerged a need for the specification of reptile parasites and their hosts.
The study aims to gain data on intestinal parasites of reptiles kept as pets or in small private collections in close contact with people.
A combination of native and iodine stained direct smears along with flotation concentration were used to investigate parasites in pet reptiles’ feces. All samples were investigated macroscopically and a smear was prepared and stained by modified Ziehl Neelsen for detection of Cryptosporidium.
Stool samples from 100 pet or small zoological reptile collections (Lacertilia=36, Serpentes=20, Chelonii=11, Corocodilia=1) were collected. The total occurrence of parasite was 52%. 64.8% of the examined Lacertilia, 35.3% of Serpentes, 45.5% of Chelonii were infected. Eimeria, Isospora, Cryptosporidium, Trichomonas, Balantidium, Strongylid and Oxyurid eggs and amoeba were identified. Cryptosporidium was detected in Lacertilita, Serpentes and Chelonii. In the only sample from a Nile crocodile no parasites were detected. Eimeria was detected in Bearded dragon, Indian python, Albino python and king cobra and Isospora was identified in Bearded dragon and the alien Cheloniid species Red-eared slider. Amoeba was identified in Iguana iguana and Horsfield tortoise.
Trichomonads, Balantidium, Cryptosporidium, Isospora, Eimeria, amoebae and nematode eggs were identified in the investigated samples. Cryptosporidium were detected by specific stains in 14 samples. Sauria was the most infected suborder (64.8%) while 32.4% of snakes and 45.5% of chelonians were infected. Parasites are common in pet reptiles but the parasite species, the degree of infestation and hygienic management will determine the ultimate clinical outcome of the existing parasite infections. Hence examination for endoparasites should be recommended for checking the health status of all captive or newly entering reptiles.

Rhipicephalus sanguineus is the most widely distributed tick in the world, which is partly due to its biological flexibility and the global distribution of its major host, the domestic dog. In Mediterranean region it could be principal reservoir host for Leishmania infantum, usually transmitted by the phlebotomine sand flies. In this study, we evaluated the vector potential of R. sanguineus in transmitting L. infantum to uninfected dogs.
During 2014, five dogs with clinical manifestations of canine visceral leishmaniasis (CVL), high anti-Leishmania antibody titers and tick infestation, were selected from CVL endemic areas (Tehran and Alborz provinces). At least, twenty live ticks were removed from each infected dog. After morphological identification, the ticks were divided into two groups; ticks belonging to the first group were dissected for parasitological examinations and semi-nested PCR assay, and those of the second group were selected for the transmission of CVL caused by L. infantum to uninfected dogs. Following tick infestation, all uninfected dogs were kept for 9 months and examined monthly for clinical and serological tests.
Nearly, 67% of ticks were infected by L. infantum using the semi-nested PCR. All other parasitological tests of ticks were negative. Clinical examinations and serological tests of the investigated dogs revealed negative results. Nested-PCR test results performed on splenic biopsy samples of dogs were also negative.
L. infantum-positive R. sanguineus ticks were unable to transfer L. infantum from infected dogs to healthy ones. The detection of L. infantum DNA in ticks collected from naturally infected dogs by semi-nested PCR does not prove their vectorial competence.

Chronic bee paralysis virus (CBPV) is an unclassified polymorphic single-stranded RNA virus. Among the viruses infecting honeybees, CBPV is known to induce significant losses in honeybee colonies. In this study, a total number of eighty-nine suspected apiaries from four regions of Iran (including Mazandaran, Khorasan Razavi, Hormozgan, and Kurdistan) were sampled and submitted for molecular identification. Three positive samples were detected by RT-PCR. All positive samples were confirmed by sequencing. The phylogenetic tree which displays the molecular relationship between the viruses of different Iranian geographic regions and references isolates was constructed. The Iranian isolates formed two distinct phylogenetic groups (Group 1 and Group 2). The IR-CPV-GMG-1, IR-CPV-GMG-2, IR-CPV-GMG-4, and IR-CPV-GMG-6 formed Group 1 and IR-CPV-GMG-3, IR-CPV-GMG-5, and IR-CPV-GMG-7 were in Group 2 as a distinct group. Iranian isolates in group 1 were similar to European and East Asian CBPVs. This research was the first phylogenetic analysis of CBPV in Iran. Further researches are needed to study the other aspects of this virus-like genetic characteristics and pathogenesis in Iran.

Biological control of parasites by using entomopathogen fungi is the one of the recommended ways to control them instead of using the chemical agents. Entomopathogen fungi are not pathogenic for animals and plants, while ticks are one of the most important parasites of animals that can transmit very important microbial pathogens. Ixodes ricinus is a hard tick that infests animals and human.
This study demonstrated enzyme assay of entomopathogen fungi hosted on Ixodes ricinus.
Enzymatic activities of chitinase, lipase and protease of fungal structures on the killed tick bodies have been assayed by standard sphectrophotometric methods.
Chitinase, lipase and protease activities showed significant differences among different fungal strains (p This study reveals the relationship between enzyme level of fungal strains and the possibility of selecting more effective strains of entomopathogenic fungi.

Rhipicephalus (Boophilus) annulatus tick is one of the most important ectoparasite of cattle. Re­cently, several laboratories in the world have been concentrated on immunizing cattle against tick using various types of tissue extracts of ticks. The aim of this study was to evaluate the effect of immunization of cattle with tick salivary gland extract on biological parameters of ticks and humoral immune responses of cattle. Fourteen more dominant protein bands identified as immunogenic by Western-blot analysis were eluted from polyacrylamide gel. Test and control groups were injected three times with eluted proteins and sterile PBS (pH= 7.2) respectively with equivalent amount of adjuvant. After four weeks a tick challenge was performed. Fi­nally, biological parameters of collected engorged female ticks were recorded and humoral immune responses to immunization measured by ELISA. The results indicated immunization of cattle resulted in reduction in mean tick counts, attachment, en­gorgement weights, feeding index, egg mass weight, hatchability and fertility index (respectively 63.1%, 62.6%, 30.2%, 36.4%, 40%, 78.7% and 13.3%) and increased duration of feeding, pre-oviposition and incubation period of eggs (respectively 8.6%, 45 and 31.34%). All changes were statistically significant (P< 0.05). Results showed an increase in antibody production of test group from the first week after immunization. The antibody level was boosted following tick infestation. This investigation indicates that immunization of cattle with these antigens could induce a protective immune response against Rh. (B.) annulatus tick that would be expected to provide a safe non-chemical means of tick control.

Despite the use of prophylactic chemotherapy and vaccination, coccidiosis is still one of the most devastating diseases in poultry industry. Understanding the immune mechanism helps researchers to prevent this parasitic infection more effectively.

The purpose of this study was to evaluate the antibody response in chickens, induced by a live attenuated vaccine (Livacox Q), before and after challenge, by means of ELISA.

One hundred and twenty one-day-old male Ross 308 broiler chicks were randomly divided into 4 groups of 30 birds. In 4th day of age, half the birds were orally vaccinated. The challenged groups received the infective dose at 14th day of age via oral administration. Besides recording weight gain, lesion score and oocyst count in 21st day old birds, humoral immunity was assessed by means of ELISA on serum samples taken from 7 and 21day-old birds.

Three days post vaccination, the average of optical density (OD) showed significant difference between vaccinated (0.553) and unvaccinated (0.686) birds (p≤0.05). In 21 day-old birds, the OD of unvaccinated-unchallenged (negative control) groups (0.331) differed significantly with vaccinated-unchallenged (0.663) birds. The average of lesion score in vaccinated-challenged birds (2.22) showed significant dissimilarity with unvaccinated-challenged groups. No difference and correlation were observed in comparing average of weight gain and oocyst count with serum optical density among treatment and control groups.

The results indicated that ELISA can be used for evaluating immunity uniformity of a flock after vaccination. Besides inducing antibody responses comparable to challenge with wild oocysts, vaccination with live attenuated coccidiosis vaccines may have inhibitory effects in intestinal lesion scores which are responsible for pathogenesis and economic loss during coccidial infections.