Boophilus Annulatus Research Articles

Effect of high concentrations of lufenuron, pyriproxyfen and hydroprene on Rhipicephalus (Boophilus) annulatus

The activity of high doses of three insect growth regulators (IGRs), lufenuron (MATCH®), pyriproxyfen® and hydroprene (Gentrol®), were tested on Rhipicephalus(Boophilus) annulatus adult females, eggs and larvae. Different concentrations of the IGRs were tested on eggs, larvae and adult ticks through immersion, larval packet and adult immersion bioassays, respectively. The tested IGRs did not show adulticidal activity against female ticks even at very high concentration. However, both hydroprene and pyriproxyfen caused a significant decrease (P<0.05) in the reproductive indices of adult female ticks. Both lufenuron and pyriproxefen showed considerable ovicidal activity delaying the hatchability of the treated eggs until the 21st day and decreasing the hatchability percentages to 37.7% and 60.6% at concentrations ≥10X and ≥4X, respectively. Lufenuron (≥10X dose), hydroprene (≥4X dose) and pyriproxyfen (≥4X dose) induced highly significant larvicidal activity as they caused 100% mortality after 72 h of exposure. The oxidative profile of the hydroprene treated ticks had decreased glutathione peroxidase and increased malonaldehyde in comparison to the other IGR- treated and control untreated ticks. It is concluded that the IGRs did not show R. annulatus adulticidal effect, however, the deposited egg mass and its hatching percent decreased significantly when treated with hydroprene and pyriproxyfen. The tested IGRs showed larvicidal activity against R. (B.) annulatus.

The acaricidal effect of peracetic acid against Boophilus annulatus and Argas persicus

The acaricidal activity of peracetic acid, PAA, against the cattle tick, Boophilus annulatus, and the fowl tick, Argas persicus was tested using filter paper contact bioassay. PAA had a great potential as acaricide and its effectiveness increased with increasing concentration and exposure time. Two minutes after treatment with 0.5%, PAA induced 100% mortality of both tick species and LC50 values for cattle and fowl ticks, after treatment for 30 min, were 0.06 and 0.05%, respectively. Followin g treatment with 0.25%, the LT50 values were 0.02 and 3.12 min, respectively. Furthermore, the detrimental effect of PAA against cattle tick extended beyond the adult stage, it significantly prolonged the preoviposition period, shortened the oviposition period, and decreased the mean number of the layed eggs, such parameters were 14.75 and 6.57 days, as well as 457.50, respectively, after treatment with 0.25% of PAA. Therefore, PAA is highly effective when used at lower doses and short exposure time. The high speed of killing ticks is very important for avoidance of the hazard ensued by pathogen transmission in the course of delayed mortality caused by the currently used acaricides.

In vitro acaricidal effects of ethanolic extract and its fractions of Ageratum conyzoides L. against common cattle tick, Rhipicephalus (Boophilus) annulatus

In vitro acaricidal effects of ethanolic extract and its fractions of Ageratum conyzoides L. against common cattle tick, Rhipicephalus (Boophilus) annulatus

Ultrastructural analysis of oocytes of Rhipicephalus (Boophilus) annulatus during postengorgement period as a tool to evaluate the cytotoxic effects of amitraz and deltamethrin on the germinative cells

The present study utilizes the ultrastructural analysis of the fully engorged female Rhipicephalus (Boophilus) annulatus ticks, as a tool to evaluate the cytotoxic potential of deltamethrin and amitraz on the germinative cells. The ultrastructural analysis of the ovary of the normal (untreated) R (B.) annulatus revealed, oocytes in different stages of development, attached to the ovary wall by pedicel cells. The attachment site of oocyte to the pedicel cell was characterized by indentations of the plasma membrane. The oocyte was bound by three cell membranes viz., plasma membrane, chorion and basal lamina. The stages of oocytes were differentiated ultrastructurally based on the features of their outer membrane and the number and size of lipid and yolk droplets. Detailed day wise analysis of ultrastructural changes in the ovary during the post-engorgement period revealed the occurrence of the degenerative changes from day five onwards. These appeared first in the oocytes followed by the germinal epithelium. The ovary of ticks treated with methanol (control), revealed similar topographies as that of a normal ovary except for the presence of very few oocytes with ring shaped nucleoli. Ultrastructurally, treatment with deltamethrin produced more prominent and extensive morphological alterations when compared to amitraz. In the case of ticks treated with amitraz, the oocytes of stage IV and V showed wavy and disrupted outer boundaries along with the loss of integrity of the yolk droplets. Uneven nuclear membranes of stage II oocytes and cristolysis of mitochondria of mature oocytes were the other changes noticed. Ticks treated with deltamethrin revealed prominent modifications such as, detachment of the basal lamina, wrinkled boundary, inconsistent nuclear membrane, ring shaped nucleoli and chromatin clumping in the case of the early stage oocytes (I and II), whereas swelling and cristolysis of mitochondria were seen in mature oocytes. The study further indicated that, in addition to the previous proven neurotoxic effects, these compounds act directly on the ovary of tick.

BIOCHEMICAL, PATHOLOGICAL AND MOLECULAR STUDIES ON BABESIOSIS IN CALVES

Bovine babesiosis is caused by the tick-borne blood parasite, Babesia bovis (B.bovis) and Babesia bigemina (B.bigemina). Many studies from 1980’s to 2015’s revealed that annual incidence of disease in summer season especially in adult cattle has associated with considerable economic losses. The present study were made on 115 Holstein calves at period from November 2014 till January 2015. Clinically affected calves showed signs of babesiosis as fever (41c°), anorexia, depression, weakness, pale mucous membrane, emaciation, weight loss, hematuria with accelerated heart and respiratory rates. Parasitological examination using Giemsa-stained thin blood films revealed presence of B. bovis in 54 out of 115 of examined calves while molecular examination by using regular Polymerase Chain Reaction (PCR) technique for same calves showed positive cases in 66 out of 115 and 14 out of 20 groups, Boophilus annulatus (B. annulatus) samples that affect these calves. There was a significant decrease in erythrocytic count, hemoglobin content, packed cell volume, albumin, globulin and total protein but significant increase in serum bilirubine in clinically affected cases of Babesia bovis. Pathological examination of liver and spleen showed golden yellow to brown hemosiderine pigment with inflammatory cells infiltration. In conclusion, the present study indicates that the change of some epidemiological aspects of clinical disease as age and season could be occure. Infected cases can occur at any time of the year in young calves less than three months. It could be occure in winter season, in contrast to some previous studies.

Molecular characterization of Babesia and Theileria species in ticks collected in the outskirt of Monte Romano, Lazio Region, Central Italy.

In 2012-2013, an investigation was carried out in the Viterbo province, Lazio region, on ticks and tick-borne Apicomplexan protozoa of the Babesia and Theileria genera. This followed the reporting of high density of ticks by soldiers operating in a military shooting range, and the signaling by owners and local veterinary authorities of several cases of babesiosis among cattle. A total of 422 ticks were collected from 35 heads, whereas 96 ticks were collected by dragging. Ticks were identified as Rhipicephalus (Boophilus) annulatus Say (n = 373), Rhipicephalus bursa Canestrini & Fanzago (n = 63), Rhipicephalus sanguineus/turanicus (n = 32), Hyalomma marginatum Koch (n = 49) and Dermacentor marginatus Sulzer, 1776 (n = 1). A randomly selected sample of ticks (235 from animals and 36 by dragging) was analyzed using molecular methods to detect species of Babesia and Theileria. In total, 11 ticks collected from animals (4.7%) and two ticks (5.5%) collected by dragging were positive. Sequencing of PCR products of the small subunit ribosomal RNA gene revealed Babesia caballi (n = 2), Babesia bigemina (n = 3), Theileria sergenti/buffeli/orientalis (n = 7) and Theileria equi (n = 1). None of the detected species has been associated with human infection.

In vitro effects of caffeic acid, nortriptyline, precocene I and quercetin against Rhipicephalus annulatus (Acari: Ixodidae).

In the present study, the acaricidal effects of caffeic acid, nortriptyline, precocene I and quercetin against Rhipicephalus annulatus (syn. Boophilus annulatus) Say (Acari: Ixodidae) were evaluated. Adult immersion technique (24 ticks immersed for 2min in one dilution of the compound) was used for the assessment of the effects of caffeic acid (0.39-100mg/mL), nortriptyline (0.625-50mg/L), precocene I (0.004488-5mg/mL) and quercetin (6.25-100mg/mL) against R. annulatus. Adult tick mortality, reproductive index, inhibition of fecundity and hatching were calculated. Caffeic acid, nortriptyline, precocene I and quercetin revealed very low adult mortality and inhibition of fecundity, even at the highest concentration tested. Quercetin (>50mg/mL) caused blocking of hatching of eggs.

Comparative in Vitro Study on the Efficacy of Some Commercial Insecticides against Rhipicephalus (Boophilus) annulatus Ticks

Commercially available preparations including Butox® 50% (containing 5% deltamethrin), Diazinon 15% and Achook 0.15% (containing 0.15% Azadirachtin) were compared for their acaricidal activity against Rhipicephalus (Boophilus) annulatus engorged females obtained from naturally infested cattle from Rashid city, Behera Province, Egypt, and other developmental stages using immersion test. It was found that the highest mortality (86.6%) was achieved by using Diazinon 15% and Achook at concentrations of 4 ml/l and 0.15%, respectively. The complete inhibition of oviposition was observed when using Butox® 50% and diazinon 15% at all used dilutions while Achook caused drop in reproductive index (RI) of tested females reached 75% at concentration of 0.075%. On the other hand, the bioassays on eggs and larvae revealed the superior effect of Achook over Butox® 50% and Diazinon 15%; as the lowest hatchability (0%) of eggs and highest mortality (100%) within the first 24 hours post treatment on unfed larvae was achieved in Achook at both concentrations of 0.15% and 0.075% azadirachtin. Finally, it was concluded that Butox® 50%, Diazinon 15% and Achook 0.15% commercially available pesticides showed variation in percentages of adult tick and larval mortalities, inhibition of oviposition and hatching of laid eggs. Promising results of Achook which is a product for agricultural use and does not have recommended dose for veterinary application yet needs further investigations to be recommended.

Effects of Ethanolic Extract of the Leaves of Pongamia glabra and Gliricidia sepium against Rhipicephalus (Boophilus) annulatus

Effects of Ethanolic Extract of the Leaves of Pongamia glabra and Gliricidia sepium against Rhipicephalus (Boophilus) annulatus

Tick (Acari: Ixodidae) infestations in cattle along Geba River basin in Guinea-Bissau

Tick infestations are a major problem for animal production in tropical areas where prevention and control remain deficient. The present study sought to assess the awareness of traditional cattle producers towards the importance of ticks and aimed at the identification of tick species infesting bovines within the Geba River basin, Guinea-Bissau. Interviews with producers revealed that the majority directly correlates the presence of ticks with the occurrence of diseases in cattle. However, insufficient or inadequate control approaches prevail. A total of 337 ticks were collected on bovines at 18 different villages (10 during dry season, and 8 during rainy season). The tick species collected during the dry season were Rhipicephalus (Boophilus) geigyi (56.5%), followed by Amblyomma variegatum (23.3%), Rhipicephalus (Boophilus) annulatus (17.6%) and Hyalomma truncatum (1%). In the rainy season A. variegatum was the most collected (88.9%), followed by R. (Boophilus) geigyi (4.2%), R. (Boophilus) annulatus (3.4%), Rhipicephalus sanguineus group (2.8%) and H. truncatum (0.7%). To support species identification, segments of both cytochrome c oxidase I (COI) and 12S ribosomal RNA (12S) genes were sequenced and the data gathered were analysed by maximum likelihood and parsimony. Morphological and genetic data of individual specimens gathered in this study provide relevant information for future studies on tick population dynamics in the region. In addition, it led to a deeper characterization of R. sulcatus and a R. sanguineus-like specimen, exploring their genetic relationship with other R. sanguineus, which supports their classification as distinct species within R. sanguineus group.

Pyrethroid resistance in Iranian field populations of Rhipicephalus (Boophilus) annulatus

Resistance to acaricides in ticks is becoming increasingly widespread throughout the world; therefore, tick control requires resistance monitoring for each tick species. The aims of this study were to monitor the susceptibility status of the cattle tick, Rhipicephalus (Boophilus) annulatus (Acari: Ixodidae), against pyrethroid acaricides from Mazandaran Province, northern Iran, and where resistance was evident, and establish the possible underlying mechanisms. Fully engorged adult R. (B.) annulatus females collected on cattle from Mazandaran Province. Twenty-nine tick populations produced 10–18days old larvae and bioassayed with cypermethrin and λ-cyhalothrin by larval packet test and the levels of detoxification enzymes were measured. Population AM-29 had a maximum resistance ratio (RR99) of 20.21 to cypermethrin and 53.57% of the tick populations were resistant at LC99 level. With λ-cyhalothrin, 17.86% of the tick populations were resistant and AM-29 was the most resistant population with RR99=4.54. AM-29 also showed significant elevation of cytochrome P450 monooxygenase (P450) and glutathione S-transferase (GST) (2.76- and 2.39-fold, respectively) (P<0.001). Rhipicephalus (Boophilus) annulatus showed resistance to pyrethroid insecticides with elevated levels of P450, GST and para-nitrophenyl acetate (p-NPA) in resistant populations. Operational failure was noted in controlling R. (B.) annulatus by pyrethroid insecticides, therefore alternative pest management measures should be adopted in Iran. For the first time, a new estimate of insecticide resistance based on effective dose recommended by the pesticide manufacturer termed Operational Dose Ratio (ODR) is defined and discussed.

Clinical and Diagnostic Studies with Special Reference to Multiplex PCR for Detection of Babesia Bovis and Anaplasma Marginale from Imported Beef Cattle in Egypt

The present study aimed to diagnose mixed infection with both Babesia. Bovis ( B. bovis) and Anaplasma marginale (A. marginale) in imported Uruganian Frisian beef cattle that transported from Alexandria to Ismailia governorates-Egypt using multiplex PCR. Twenty five blood samples from investigated beef cattle were collected and examined using different parasitological, heamatological and molecular investigations. Ticks infesting the examined animals were collected and tested for presence of babesia and anaplasma by molecular technique. Clinically infected beef cattle showed variable clinical signs as pyrexia up to 40 - 41°C, anorexia, dullness, depression, weakness, pale oricteric conjunctival mucous membranes, dyspnea, accelerated respiratory and heart rates, coughing, harsh vesicular sound, rattling tracheal sound, oculo-nasal discharge, petechial hemorrhage of buccal and conjunctival mucus membrane, bloody feces and nervous manifestation including ataxia and in coordination. Four beef cattle were found dead without apparent clinical signs. Hematological results showed a significant decrease in erythrocytic count, hemoglobin content, and packed cell volume in clinically affected cases. Parasitological examination using Giemsa-stained thin blood smearsrevealed presence of a mixed infection with B. bovis and A. marginale in 8/25 (32%) and A. marginale alone in 10/25 (40%).Molecular examination using PCR assay showed that mixed B. bovis and A. marginale were detected on 10 samples (40%) and 12 samples (48%) were positive for A. marginale alone while one sample (4%) was positive for B. bovis alone and the last two samples (8%) were completely negative. Out of five ticks Boophilus annulatus (Bo. annulatus) groups, three (60%) were positive to mixed infection and 2(40%) for A. marginale alone. It was recommended to tighten the quarantine measures and rules to avoid introduction of expatriate diseases and protect the newly imported animals.

First report of pyrethroid resistance in Rhipicephalus (Boophilus) annulatus larvae (Say, 1821) from Iran

Rhipicephalus (Boophilus) annulatus is one of the most important hard ticks parasitizing cattle in northern Iran. The aim of this study was to evaluate pyrethroid resistance levels of this species from Nur County, northern Iran. The hard ticks were collected through a multistage cluster randomized sampling method from the study area and fully engorged female R. (B.) annulatus were reared in a controlled insectary until they produced larvae for bioassay. Seventeen populations of the hard ticks were bioassayed with cypermethrin and 12 populations with lambda-cyhalothrin using a modified larval packet test (LPT). Biochemical assays to measure the contents/activity of different enzyme groups including mixed function oxidases (MFOs), glutathione S-transferases (GSTs) and general esterases were performed. Population 75 showed a resistance ratio of 4.05 with cypermethrin when compared with the most susceptible field population 66 at the LC50 level. With lambda-cyhalothrin the resistance ratio based on LC50 was 3.67 when compared with the susceptible population. The results of biochemical assays demonstrated significantly elevated levels of GSTs and esterases in populations tested compared with the heterozygous susceptible filed population and a correlation coefficient of these enzymes was found in association to lambda-cyhalothrin resistance. Based on the results, pyrethroid acaricides may operationally fail to control R. (B.) annulatus in North of Iran. This study is the first document of pyrethroid resistance in R. (B.) annulatus populations from Iran.

Identification and Infestation of Ticks on Cattle in the Peri-Urban Area of Ngaoundere, Cameroon

Identification and Infestation of Ticks on Cattle in the Peri-Urban Area of Ngaoundere, Cameroon Livestock production yield in the sub-Saharan Africa is influenceby ticks and the diseases they transmit. The aim of this study was to identify the tick species in the peri-urban area of Ngaoundere, the capital of the Adamawa region (Cameroon), where traditional farms co-exist, lack of acaricide products usage and no modern farming. This study was conducted from August to September in 5 localities and in 8 farms. All visible ticks were collected from the animals selected for the study and preserved in 70% ethanol. Ticks collected were counted and morphologically identified. The collection of ticks showed that Tropical Bont Tick, Rhipicephalus (Boophilus) geigyi, Rhipicephalus (Boophilus) annulatus, Rhipicephalus (Boophilus) decoloratus, Hyalomma truncatum, Hyalomma marginatumrufipes, Rhipicephalus sanguineus and Haemophysalys laechi were the main tick species and sub- species found in the sub-urban area of Ngaoundere, with prevalences: 24.4%; 1.97%; 0.91%; 3.26%; 0.18%; 0.74%; 0.88%; 0.04% respectively. The use of acaricides reducedsignificantly (p<0.01) the infecting load of ticks and contributes to the improvement of the hematocrit (p<0.01). Young cattle are more likely to be harmed by ticks and tick-borne diseases, as they are more infected andanaemic than adults (p<0.05). The results of this study revealed that the services of a veterinarianis required in orderto establish a real strategy against ticks and use of acaricides to reduce the tick population hence improving cattle physiological parametersas well as productivity.

Identification and characterization of a cathepsin L-like cysteine protease from Rhipicephalus (Boophilus) annulatus.

The tick Rhipicephalus (Boophilus) annulatus is one of the most important ectoparasites of bovines and is responsible for the transmission of different pathogens such as Babesia and Anaplasma. Cysteine proteases are involved in several host-tick interactions including invasion of host tissues, immune evasion, pathogen transmission, embryogenesis and blood digestion. In this study, the gene encoding R. annulatus cathepsin L-like enzyme (RaCL1) was cloned into pTZ57R/T vector, sequenced and analyzed using bioinformatics approaches. The nucleotide length of RaCL1 was 999 bp. Bioinformatics analysis showed 332 amino acids with an approximate molecular weight of 36.3 kDa which contained a signal peptide sequence (18 amino acids), pro-region (97 amino acids) and mature enzyme (217 amino acids). Multiple sequence alignment of the RaCL1 revealed high similarity to cathepsin L-like cysteine proteases from other tick species such as Rhipicephalus (Boophilus) microplus and Amblyomma variegatum. Based on bioinformatics analyses, results of this work suggest that RaCL1 can be a suitable candidate for the development of vaccine against R. annulatus.

Isolation of aerobic bacteria from ticks infested sheep in Iraq

ObjectiveTo highlight the presence of aerobic bacteria in live ticks infested sheep, in Diyala Governorate, Iraq. MethodsOne hundred and thirty adult alive ticks were picked up from sheep which were reared in different farms in Diyala Governorate, Iraq, during the period from November 2012 to May 2013. Ticks were classified in the Natural History Museum in Baghdad. They were dissected aseptically for extraction of the salivary gland and mid-gut. The removed tissue from each organ was inoculated in buffer peptone water (1%) and incubated for 2 h at 37 °C, to maintain weak and/or injured bacterial cells, then transmitted to nutrient broth incubated at 37 °C for 18 h. Culturing was done on three solid bacteriological media (nutrient, blood and McConkey agars), and then incubated at 37 °C for 24 h. Bacterial identification was performed by using multiple biochemical tests and API-20 strips. Data were analyzed by using Statistical Analysis System version 9.1, 2010. Chi-square test was used for comparison at significant level of P ≤ 0.05. ResultsTwo species of ticks were identified [Rhipicephalus (Boophilus) annulatus and Hyalomma turanicum]. High bacterial isolation rate was observed (483 isolates). A significant high isolation rate was recorded from Rhipicephalus annulatus (63.14%). Six bacterial species were identified [Escherichia coli (28.36%), Pseudomonas aeruginosa (18.01%), Bacillus cereus (14.69%), Staphylococcus aureus (13.66%), Citrobacter freundii (13.04%), and Enterobacter species (12.21%)]. Also the high bacterial isolation rates were recorded in the temperate months (November, March and April); these coincided with high reproductive performance of ticks. ConclusionsThe high isolation rate of aerobic pathogens from ticks might reflect the active contribution of this arthropod in environmental contamination and increase the probability of transmitting bacterial pathogens to their hosts.

Contrasting effects of ethanolic extracts of leaf and flower of Chromolaena odorata against Rhipicephalus (Boophilus) annulatus

In the present study, ethanolic extracts of leaves and flowers of Chromolaena odorata were compared at different dilutions (6.2 mg/mL, 12.5 mg/mL, 25 mg/mL, 50 mg/mL and 100 mg/mL) for their efficacy against ticks. Per cent adult mortality, inhibition of fecundity and hatching of laid ova were studied. Leaf extract did not cause any adult tick mortality. The negative values for the per cent inhibition of fecundity observed with the leaf extract indicatedthat it promoted egg laying. On the contrary, the ethanolic extract of flowers at 10% concentration caused 62% adult tick mortality and 80%inhibition of fecundity. However, both extracts did not produce any change in the hatching of the laid ova by treated ticks.

Molecular characterization of Rhipicephalus (Boophilus) annulatus from Iran by sequences of cytochrome c oxidase subunit I (COI) and the second internal transcribed spacer (ITS2)

Background: Traditionally, morphological features of Rhipicephalus (Boophilus) annulatus from closely-related ticks have been considered for their identification and differentiation. However, it is difficult and requires expertise in order to accurately identify and differentiate engorged female ticks and some developmental stages such as larva and nymph from other similar ticks. Hence, molecular markers may be a suitable alternative. OBJECTIVES: Mitochondrial cytochrome c oxidase subunit I (COI) gene and the second internal transcribed spacer (ITS2) fragments of Rh. (Bo.) annulatus were sequenced to assess the use of molecular techniques for identifications and phylogenetic studies of these ticks. METHODS: Polymerase chain reaction (PCR) technique was performed based on the analyses of COI and ITS2 sequences of ticks collected from two different regions in Iran (Golestan and Mazandaran). RESULTS: The length of COI and ITS2 sequences were 1539 and 1158bp, respectively. The nucleotide similarity of COI gene was 91.3% between the ticks examined from the two different regions. The deduced amino acid sequences from COI showed 98.6% similarity between the ticks studied and showed 98.2 and 99.6% similarity with the only complete sequence of Rh. (Bo.) annulatus (AGH19677) registered in GenBank. The obtained complete nucleotide sequences of ITS2 from Rh. (Bo.) annulatus from Golestan and Mazandaran revealed 99.9% similarity, while the other ticks registered in GenBank 95 to 99% similarity (KC503267, AF271270, AF271272, JQ412126). CONCLUSIONS: It seems that COI and ITS2 sequences could provide suitable genetic markers for discrimination and genetic characterization of Rhipicephalus (Boophilus) annulatus.

Distribution of Tick Species Spread in Kütahya Vicinity

This study was performed to determine the tick species infested to the cattle and small ruminants in Kütahya Province and their seasonal activity and prevalence. Examination was performed in terms of infestation in 2402 farm animals and their shelters in 10 villages between October 2010 and October 2011. The infestation rate of animals was detected as 9.55%. A total of 657 adult ticks were collected. A total of 11 tick species consisting of Ixodes ricinus, Ixodes hexagonus, Rhipicephalus (Boophilus) annulatus, Dermacentor marginatus, Hyalomma marginatum, Haemaphysalis parva, Haemaphysalis sulcata, Haemaphysalis punctata, Rhipicephalus bursa, Rhipicephalus sanguineus and Rhipicephalus turanicus were determined. Seasonal fluctuations of tick species were also determined. Ixodes ricinus and R. (Boophilus) annulatus species were encountered mostly during fall and winter months. Dermacentor marginatus was usually found in winter time. Hyalomma marginatum, was found between April and July. Haemaphysalis parva was found in October, November and March. Haemaphysalis sulcata was found in November and March. Haemaphysalis punctata was found only in March and November. Rhipicephalus species were generally found in late spring and summer. Finally, these results will be helpful for the determination of control methods of tick infestations and tick-borne diseases in the region.

Histoarchitecture of the ovary of Rhipicephalus (Boophilus) annulatus during pre- and postengorgement period.

The present communication describes the detailed day wise study of histological changes of the ovary of Rhipicephalus (Boophilus) annulatus in the postengorgement period together with the systematic classification of their oocytes. The ovary of R. (B.) annulatus is panoistic type with an asynchronous development of oocytes. All the stages (II, III, IV, and V) of oocytes except stage I were similar to R. (B.) microplus. The stage I oocytes showed basophilia, which was not reported earlier in other species of ticks. Day wise changes were in the form of presence of oogonia in partially fed and day one engorged adults, considerable degeneration of oocytes on day two, emergence of new wave of oocytes on day three, presence of mature oocytes up to day eight, and complete degeneration of ovarian tissue from day eight onwards. The degenerative changes in the ovary appeared initially in the oocytes followed by germinal epithelium.