Development of Hyalomma lusitanicum under laboratory conditions

At the constant temperature of 25 degrees C and relative humidity (RH) of 84%, the average pre-oviposition period of Hyalomma lusitanicum was 47 days, the oviposition lasted an average of 26 days and the total egg production was an average of 6320 per female. At 16 degrees C the females did not lay eggs at all, but those which survived for 1 year and were transferred thereafter to 25 degrees C and 84% RH laid viable eggs. At 35 degrees C, the oviposition was identical at all levels of RH tested (25, 62 and 93%). At 25 degrees C, the pre-oviposition period was shorter at 93% RH, and the number of eggs laid was fewer at 25% RH. The eggs hatched in 32-40 days, the hatching percentage being lower in batches of eggs laid at the beginning and at the end of the oviposition period. The larval and nymphal moultings were not influenced by the type of host. As the temperature increased, the pre-moult period became shorter. The engorged larvae were more sensitive to the low RH than the engorged nymphs, whose moulting percentage was always greater than 72 in all regimes. Low temperature and high humidity had a favourable effect on the survival of unfed nymphs. The female-to-male ratio was 1:2. Hyalomma lusitanicum always behaved as a 3-host tick. The adults did not engorge on rabbits. The female ticks engorged on calves weighed an average 543 mg. Ticks maintained at 25 degrees C and 84% RH and engorged on calves completed the life cycle in 138-196 days, which does not include the period of chitinization of about 30 days. More than half of this period was spent in egg laying and hatching.     

Effect of temperature on transovarial transmission of Babesia bigemina (Smith and Kilborne, 1893) in Boophilus annulatus (Say, 1821)

The effect of the temperature on the transovarial passage of Babesia bigemina has been studied in female Boophilus annulatus during their oviposition. Kinetes of Babesia were present in eggs laid at the temperatures of 16, 20, 26, 28, 30 and 35 degrees C. The rate of infection was temperature dependent. It reached at least 50%, even at low temperatures. The average infection level at 26 and 30 degrees C was 270 kinetes per egg. However, between 20 and 35 degrees C the eggs laid during the first 3 days were parasite free. At 16 degrees C, no kinetes were detected during the first 13 days of oviposition.     

Transmission of Babesia spp by the cattle tick (Boophilus microplus) to cattle treated with injectable or pour-on formulations of ivermectin and moxidectin

OBJECTIVE: To assess the efficacy of ivermectin and moxidectin to prevent transmission of Babesia bovis and Babesia bigemina by Boophilus microplus to cattle under conditions of relatively intense experimental challenge. DESIGN: Naive Bos taurus calves were treated with either pour-on or injectable formulations of either ivermectin or moxidectin and then exposed to larvae of B microplus infected with B bovis or larvae or adults of B microplus infected with B bigemina. One calf was used for each combination of haemoparasite, B microplus life stage, drug and application route. PROCEDURE: Groups of calves were treated with the test drugs in either pour-on or injectable formulation and then infested with B microplus larvae infected with B bovis or B bigemina. B bigemina infected adult male ticks grown on an untreated calf were later transferred to a fourth group of animals. Infections were monitored via peripheral blood smears to determine haemoparasite transmission. RESULTS: Cattle treated with either pour-on or injectable formulations of ivermectin and moxidectin became infected with B bovis after infestation with infected larvae. Similarly, larvae infected with B bigemina survived to the nymphal stage to transmit the haemoparasite to animals treated with each drug preparation. Cattle treated with pour-on formulations of ivermectin and moxidectin then infested with adult male ticks infected with B bigemina did not become infected with B bigemina whereas those treated with the injectable formulations of ivermectin and moxidectin did show a parasitaemia. CONCLUSIONS: Injectable or pour-on formulations of ivermectin and moxidectin do not prevent transmission of Babesia to cattle by B microplus. Use of these drugs can therefore not be recommended as a primary means of protecting susceptible cattle from the risk of Babesia infection.     

Babesia spp. infection in Boophilus microplus engorged females and eggs in Sao Paulo State, Brazil

Babesia spp. infections were investigated in Bos taurus x Bos indicus dairy cows and calves and in Boophilus microplus engorged female ticks and eggs. Blood samples and engorged female ticks were collected from 25 cows and 27 calves. Babesia spp. was detected in ticks by microscopic examination of hemolymph of engorged female and by squashes of egg samples. Cattle infection was investigated in blood thin smears and by DNA amplification methods (PCR and nested PCR), using specific primers for Babesia bovis and Babesia bigemina. Merozoites of B. bovis (3 animals) and B. bigemina (12 animals) were detected exclusively in blood smears of calves. DNA amplification methods revealed that the frequency of B. bigemina infection in calves (92.6%) and in cows (84%) and of B. bovis in calves (85.2%) and in cows (100%) did not differ significantly (P > 0.05). Babesia spp. infection was more frequent in female ticks and eggs collected from calves (P < 0.01) than from cows, especially in those which had patent parasitemia. Hatching rates of B. microplus larvae were assessed according to the origin of engorged females, parasitemia of the vertebrate host, frequency and intensity of infection in engorged female tick, and frequency of egg infection. Hatching rate was lower in samples collected from calves (P < 0.01) than from cows, and in those in which Babesia spp. was detected in egg samples (P < 0.01).     

Detection of Babesia bigemina infection in strains of Rhipicephalus (Boophilus) microplus collected from outbreaks in south Texas

The sudden death of several cattle infested experimentally with Rhipicephalus (Boophilus) microplus led to a clinical investigation into the reasons for the unexpected mortality. Microscopic evidence for Babesia bigemina infection was found in blood smears from the affected animals and a PCR assay was designed to detect the presence of B. bigemina and Babesia bovis in all R. microplus strains received and propagated at the laboratory. The assay utilizes a nested PCR approach with the first PCR amplifying a well-conserved segment from the Babesia 18S ribosomal RNA gene followed by a nested PCR with Babesia species-specific primers and annealing temperatures enabling amplification of the 18S ribosomal RNA gene fragment specific to either B. bigemina or B. bovis. DNA from groups of 50 larvae was extracted using a rapid DNA preparation protocol, which consisted of grinding the frozen tick larvae in PCR buffer and boiling the mixture for 5min. The assay sensitivity allowed for the detection of the equivalent of a single infected tick larva. R. microplus eggs were also analyzed, but yolk protein viscosity created inconsistent results with the crush and boil DNA isolation protocol, necessitating the use of a more extensive proteinase K digestion-based DNA purification method. We detected the presence of B. bigemina in all strains of R. microplus currently reared at the laboratory and 4 of 26 strains collected from infestation outbreaks in Texas by the U.S. Cattle Fever Tick Eradication Program.     

Effect of temperature on development of the free-living stages of Ostertagia circumcincta

Hatching of the eggs of Ostertagia circumcincta was studied by recovering them from faeces and incubating them in distilled water at temperatures of 4, 16, 25 and 35 degrees C. Hatching occurred at all the temperatures. The rate of hatching increased with the rise in temperature. Development of larvae to the infective third stage (L3) was studied in faecal cultures incubated at 4, 16, 25, 30, 35, 40 and 45 degrees C. Except at 4 degrees C, L3 developed at all temperatures, the optimum temperature being 16 degrees C. The rate of development of L3 increased with the rise in temperature. This resulted in a corresponding decrease in the percentage recovery of larvae.     

野蚕越冬卵的孵化和年发生代数的研究

研究证实(1980—1984年),嘉湖平原野蚕年发生三代.越冬卵始孵期为四月中旬,盛期为六月中旬.但近半个世纪以来.很多人一直误认为四月中旬发生的野蚕是第一代,六月下旬发生的为“第二代”,事实上都是越冬卵孵化的幼虫.均为第一代.经测定,四月下旬越冬卵孵化率为0.15%,5月下旬为7.9%,6日20日为91%.本区野蚕产越冬卵最的早在9月17日,高峰期在11月上旬.结束期在12月14日,越冬卵产出越早,翌年卵孵期也早,孵化的早和迟之间可相差122天.野蚕越冬卵翌年孵化期长的原因.与秋冬产卵期长及桑园生态环境复杂有关.据室内4月24日越冬卵孵化的幼虫连续饲养,到第三代卵期恰与室外第二代卵期相重叠,说明自然界发生第四代是可能的,但真正存活率在1%以下.在防治措施上,以在越冬卵孵化高峰期(6月15—20日)施用0.1%敌敌畏.防效最好.     

Characterization of a repetitive DNA probe for Babesia bigemina

A plasmid (p16) containing a Babesia bigemina DNA insert was selected and labeled with 32P. This probe was evaluated for specificity and sensitivity by dot blot hybridization. The probe was specific and hybridized with only Babesia bigemina DNA, and not DNA from Babesia bovis, bovine leukocyte, Trypanosoma brucei or Anaplasma marginale. The DNA probe detected as little as 10 pg of Babesia bigemina DNA. The probe hybridized with Babesia bigemina isolates from Mexico, the Caribbean region and Kenya. Genomic Babesia bigemina DNA of a Kenyan isolate was digested with restriction endonucleases, and the fragments were separated by gel electrophoresis and Southern blotted. The filter was hybridized with labeled p16 and each endonuclease digestion produced at least 16 resolvable DNA fragments. The inserted Babesia bigemina DNA was approximately 6.3 kb in size. A partial restriction map was constructed. A simple whole blood dot blot procedure was utilized to evaluate the sensitivity of the DNA probe. This probe would detect as few as 150 Babesia bigemina infected erythrocytes contained in a 1-microliter sample. The DNA probe has the potential to be a very sensitive and specific diagnostic tool.     

Clinical effects, virological and serological responses in chickens following in-ovo inoculation of reticuloendotheliosis virus

Six-day-old embryonated specific pathogen free chicken eggs were inoculated with reticuloendotheliosis virus (REV) into the yolk sac and were incubated until they hatched. The hatchability of eggs inoculated with REV was significantly less (P less than 0.025) than that of media-inoculated controls. Although there were no significant differences in the body weights of these chickens at hatching, there were differences (P less than 0.001) at 6, 25 and 51 days of age between the infected and control chickens. Six of 10 chickens hatched from eggs inoculated with REV had feathering defects at 6 days of age. All chickens hatched from infected eggs had cell-free viraemia and antigenaemia, but not precipitating antibodies. Some of these chickens had very low neutralising antibody titres (less than 45) when examined at 25 and 37 days of age, as did all 10 chickens at 51 days of age. A low rate of horizontal transmission was indicated by the detection of antibodies at 37 and 51 days of age in chickens running in contact with the chickens hatched from eggs inoculated with REV.     

双芽巴贝斯虫体外培养技术初探

应用双芽巴贝斯虫染虫血和液氮保藏株,进行体外培养技术初步研究,获得了生长、增殖和保种效果。双芽巴贝斯虫在牛红细胞内带虫率达５％,低染虫率的连续培养已超过２５ｄ。奠定了双芽巴贝斯虫培养技术应用基础。     

Studies on ticks of veterinary importance in Nigeria. VI. Comparisons of oviposition and the hatching of eggs of Hyalomma species

Fully engorged Hyalomma spp. in Nigeria oviposited greater numbers of eggs than those partially engorged. Hyalomma impressum was a more prolific egg layer than H. rufipes, H. impeltatum and H. truncatum. The variations in the egg output as well as the recognizable peaks in the number of eggs during oviposition were described for each species. No species of Hyalomma below the engorged weight of 0.2 g oviposited; oviposition started with ticks of weight 0.3 g. Eggs produced by ticks weighing below 0.3 g did not hatch; the highest percent egg eclosion occurred with ticks of weight 0.6 g (H. rufipes) and 0.7 g (other Hyalomma species). The pre-oviposition, oviposition and eclosion periods were shortened when eggs were laid and incubated at high temperatures, although the number of oviposited eggs did not increase significantly. At the standard temperature of 24 degrees C, the longest eclosion period was seen in the eggs of H. rufipes (41 days) while those of H. truncatum, H. impressum and H. impeltatum were similar to each other (29 days). Only eggs of H. rufipes hatched at an incubation temperature of 15 degrees C. Eggs of Hyalomma species laid at the same time hatched over a 2--4 day period, except at 15 degrees C when the hatching period of H. rufipes lasted 10 days. The eclosion period was longest in the earlier ovipositions and shorter in the later ones. It is suggested that some intrauterine larval development might have started in the eggs before they were released at a later oviposition period. The percentage mortality of eggs at various temperatures showed that eggs of H. rufipes were more tolerant of low temperatures than those of H. impressum, H. truncatum and H. impeltatum, while the eggs of the latter 3 species were more tolerant of high temperatures than those of H. rufipes. The relevance of these results of the distribution and abundance of the Hyalomma species in Nigeria was discussed.     

Babesia bovis and B. bigemina DNA detected in cattle and ticks from Zimbabwe by polymerase chain reaction

From blood collected from 94 cattle at 12 locations in the eastern and northeastern areas of Zimbabwe, DNA was extracted and analysed by polymerase chain reaction with primers previously reported to be specific for Babesia bigemina and Babesia borvis. Overall, DNA of Babesia bigemina was detected in the blood of 33/94 (35%) cattle and DNA from B. bovis was detected in 27/58 (47%) of cattle. The prevalence of DNA of B. bigemina was significantly higher in young animals (<2 years) (23/46) than in animals over 2 years of age (10/48; chi2= 8.77; P <0.01%). Although tick sampling was not thorough, Boophilus decoloratus could be collected at 7/9 sites sampled and Boophilus microplus at 4/9 sites. Of the 20 B. decoloratus allowed to oviposit before PCR analysis, 1 (5%) contained DNA that could be amplified with primers for B. bigemina while 12 (60%) were positive with primers for B. bovis. Of the B. microplus allowed to oviposit, 11/16 (69%) were positive for B. bovis DNA by PCR and 2/16 (12%) were positive for B. bigemina.     

Effect of pre-incubation storage conditions on hatchability,chick weight at hatch and hatching time in broiler breeders

1. Eggs were stored for two different times at varying temperatures. The effects on hatchability, chick weight at hatch and hatching time were examined in two broiler breeder lines from 33 to 58 weeks of age. 2. Short storage (1 to 3 d). Storage at 20 degrees C compared with 16.5 degrees C reduced hatchability of all eggs set. No effect was observed on hatchability of fertile eggs, hatching time or chick weight. 3. Long storage (9 to 11 d). Storage at 16.5 degrees C compared with 10 degrees C decreased both hatchability of fertile eggs and chick weight at hatch. Incidence of early embryonic death increased and incubation time decreased at 16.5 degrees C compared with 10 degrees C. 4. Chicks from morning eggs were heavier than those from afternoon eggs irrespective of storage conditions. 5. Hatchability (all eggs set and fertile eggs) and chick weight varied with hen age irrespective of storage conditions. During long storage, hatching time varied with hen age independently of breeder line, storage temperature or egg laying time. 6. Hatchability (all eggs set and fertile eggs) was higher in line A than in line B. Line B eggs hatched later and produced heavier chicks than line A eggs irrespective of storage time.     

Effects of temperature and relative humidity on development and survival of the free-living stages of Trichostrongylus colubriformis, T. rugatus and T. vitrinus

Temperatures of 4 and 10 degrees C reduced the rate of hatching of eggs and the development of the pre-infective stages of T. colubriformis and T. rugatus, but not T. vitrinus. Eggs of T. rugatus hatched at 4 degrees C, but did not develop to the infective stage. At 20 and 30 degrees C, no differences were detected between species in egg hatching or the development of infective larvae. Third-stage larvae of T. rugatus survived for significantly longer periods of time at 33 and 56% relative humidities at a temperature of 20 degrees C than did either T. colubriformis or T. vitrinus. At 30 degrees C, differences were less marked, but T. rugatus and T. colubriformis survived longer than T. vitrinus. The results are discussed in relationship to the known distribution of these species in eastern Australia.     

The effect of the ovine host parasitaemia on the development of Babesia ovis (Babes, 1892) in the tick Rhipicephalus bursa (Canestrini and Fanzago, 1877)

Batches of Rhipicephalus bursa adult ticks were fed on two lambs with 10.0% (batch 1) and 0.3% (batch 2) Babesia ovis parasitaemia, respectively. Haemolymph and eggs were checked for parasites daily after detachment, before and after appearance of B. ovis in the lamb's blood.B. ovis kinetes were found in the haemolymph and eggs earlier in the engorged ticks detached before appearance of the parasite in the host blood. Rates of haemolymph and egg infection with B. ovis as well as the percentage of infected eggs were much higher in batch 1 (10% lamb parasitaemia) than in batch 2 ticks (0.3% lamb parasitaemia). In eggs incubated at 28 degrees C the optimal period to look for kinetes seems to be days 4-9. Heavily infected ticks laid fewer less eggs within a shorter oviposition period. Pre-oviposition, pre-hatching periods and egg hatchability were not affected. Various parasitic forms are described in the haemolymph and the eggs.     

红茂草生物碱对黏虫卵的杀灭作用

为评价野生药用植物红茂草的生物碱提取物对黏虫卵的杀灭效果,取适量黏虫卵分别与含50、100、200、400、800 mg/L红茂草生物碱提取物的丙酮溶液短暂接触,观察并比较经不同浓度红茂草生物碱处理的黏虫卵孵化率及初孵幼虫存活率,并设对照处理组（CK）。结果显示,除50 mg/L处理组外,其余各处理组的卵孵化率均显著低于对照组（P〈0.05）;在设定的处理浓度范围内,所有处理组的初孵幼虫存活率均显著低于对照组（P〈0.05）,表明红茂草生物碱对黏虫卵具有显著的杀灭作用。     

PCR-based detection of the transovarial transmission of Uruguayan Babesia bovis and Babesia bigemina vaccine strains

Bovine babesiosis is responsible for serious economic losses in Uruguay. Haemovaccines play an important role in disease prevention, but concern has been raised about their use. It is feared that the attenuated Babesia bovis and Babesia bigemina vaccine strains may be transmitted by the local tick vector Boophilus microplus, and that reversion to virulence could occur. We therefore investigated the possibility that these strains could be transmitted via the transovarial route in ticks using a Babesia species-specific polymerase chain reaction (PCR) assay. DNA was extracted from the developmental stages of the tick vector that had fed on calves immunized with the haemovaccine. It was possible to detect Babesia DNA not only in adult ticks, but also in their eggs and larvae. In addition, it was shown that calves infested with larvae derived from eggs laid by ticks fed on acutely infected calves, were positive for Babesia using PCR. Caution should therefore be shown with the distribution of the haemovaccine in marginal areas. It is still advisable that suitable tick control measures be used to prevent transovarial transmission and the potential risk of attenuated Babesia reverting to virulence.     

THE DURATION OF LATENT INFECTION AND FUNCTIONAL IMMUNITY IN DROUGHTMASTER AND HEREFORD CATTLE FOLLOWING NATURAL INFECTION WITH BABESIA ARGENTINA AND BABESIA BIGEMINA

Tne Droughtmaster and 9 Hereford cattle were born in an enzootic babesiasis area and became naturally infected with Babesia argentina and B.bigemina during a 3 year period. They were then kept free of cattle ticks (Boophilus microplus) for the remainder of the experiment. Annually for the next 3 years their individual infection status with Babesia was determined by sub-inoculation of blood into splenectomised calves. At the end of this period the functional immunity of all cattle was challenged by blood inoculation of heterologous strains of B. argentina and B. bigemina. Infection with B. argentina persisted in all Herefords for 2 years and in 7 for 3 years after they had been freed of B. microplus. The number of Droughtmasters with detectable B. argentina infection progressively declined, and at the end of 3 years only 2 of 10 were still infected. No Herefords were shown to be infected with B. bigemina following 1 year's freedom from B. microplus but latent B. bigemina infection of at least 2 year's duration was demonstrated in one of the Droughtmasters. A marked degree of resistance was apparent in all cattle when they were challenged with an heterologous strain of B. argentina. There were no differences between the response to challenge of the Herefords and Droughtmasters nor between the reactions of cattle which had apparently naturally sterilised B. argentina infection and those which were still infected. The heterologous strain of B. bigemina produced parasitaemia in the majority of animals but only minimal fever and anaemia resulted with no significant differences between the breeds.     

Early development of artificially spawned southern mullet,Liza richardsonii (Smith, 1846)

The embryonic development of artificially spawned southern mullet, Liza richardsonii, eggs and the development of the larvae to 46 days are described and illustrated using drawings and photographs. The floating eggs hatched in sea water at 18–24°C after 46–60 h. Newly stripped eggs usually had more than one oil droplet (up to 16) which merged during development. Free embryos averaged 2,36 mm at hatching and had distinct yellow pigment.     

澳洲坚果新害虫--脊胸露尾甲生物学习性观察

在云南省西双版纳种植区发现为害澳洲坚果的新害虫——脊胸露尾甲,为准确掌握其生物学习性,在室内以澳洲坚果果仁为食对其进行了饲养,并对各虫态的形态、成虫交配习性、产卵习性、卵的孵化率、各虫态的存活率进行了观察。结果表明：脊胸露尾甲的发育经历卵、幼虫、蛹、成虫四个虫态,其中幼虫分为3个龄期,随着龄期的增长,幼虫体长、体宽和头壳宽度逐渐增加。成虫羽化后不会立即交配,交配前期为（4.65±0.04）d,交配时长不等,初次交配时长为（13.78±0.29）min,雄成虫存在争夺交配权的行为。雌成虫产卵前期为（8.65±0.03）d,初次产卵量为(1.94±0.06)粒,在15:00-18:00时间段的产卵量/小时最高;雌成虫对产卵位置具有选择性,背光面的着卵率极显著高于向光面。卵的孵化率随着湿度的增加而不断提高。常规饲养条件下,卵、1龄幼虫、2龄幼虫、3龄幼虫、蛹的存活率分别为(86.50±0.69)%、(83.31±2.99)%、(94.85±0.71)%、(97.09±1.46)%、(98.77±1.23)%。本研究为脊胸露尾甲的生物学及防治提供参考依据。