Efficacy of liquid spray decontaminants for inactivation of Bacillus anthracis spores on building and outdoor materials

Aims: To obtain data on the efficacy of various liquid and foam decontamination technologies to inactivate Bacillus anthracis Ames and Bacillus subtilis spores on building and outdoor materials. Methods and Results: Spores were inoculated onto test coupons and positive control coupons of nine different materials. Six different sporicidal liquids were spray‐applied to the test coupons and remained in contact for exposure times ranging from 10 to 70 min. Following decontamination, spores were recovered from the coupons and efficacy was quantified in terms of log reduction. Conclusions: The hydrogen peroxide/peracetic acid products were the most effective, followed by decontaminants utilizing hypochlorous acid chemistry. Decontamination efficacy varied by material type. Significance and Impact of the Study: The study results may be useful in the selection of technologies to decontaminate buildings and outdoor areas in the event of contamination with B. anthracis spores. These results may also facilitate selection of decontaminant liquids for the inactivation of other spore‐forming infectious disease agents.     

A quantitative TaqMan PCR assay for the detection of Mycoplasma suis

Aim: To develop a TaqMan probe‐based, highly sensitive and specific quantitative PCR (qPCR) assay for the detection and quantification of Mycoplasma suis in the blood of pigs. Methods and Results: Primers and probes specific to Myc. suis 16S rRNA gene were designed. The qPCR assay’s specificity, detection limit, intra‐ and inter‐assay variability were evaluated and its performance was compared with a Myc. suis conventional PCR assay (cPCR). Blood of two experimentally infected pigs, 40 Indiana pigs, 40 Brazilian sows and 28 peccaries were tested. The assay detected as few as ten copies of Myc. suis plasmids and was 100‐fold more sensitive than the cPCR. No cross‐reactivity with nontarget pig mycoplasmas was observed. An average of 1·62 × 10¹¹ and 2·75 × 10⁸ target copies ml^?1 of blood were detected in the acutely and chronically infected pigs, respectively. Three (7·5%) pigs and 32 (80·0%) sows were positive while all peccaries were negative for Myc. suis. Conclusion: The developed qPCR assay is highly sensitive and specific for Myc. suis detection and quantification. Significance and Impact of the Study: TaqMan qPCR is an accurate and quick test for detection of Myc. suis infected pigs, which can be used on varied instrumentation platforms.     

Laboratory evaluation of large-scale decontamination approaches

Aims: To evaluate the effectiveness of two spray‐based decontamination methods for surface contamination reduction and to determine the potential for contamination spread by these methods. Methods and Results: Material coupons (treated plywood and concrete) were contaminated with c. 1 × 10⁷ spores of Bacillus atrophaeus by aerosol deposition. Decontaminants (pH‐adjusted bleach or Spor‐Klenz^® RTU) were applied to coupons by either backpack sprayer or gas‐powered sprayer. Contact time, reapplication frequency and rinse method were also varied. In addition to surface removal efficacy, partitioning of contamination between the rinsate and aerosol fractions was determined. Results indicated that pH‐adjusted bleach was effective (≥6 logs reduction) when two applications and a 30 min contact time were administered, regardless of the decontaminant application method or material. Spor‐Klenz^® RTU was effective on wood, but achieved ≤3 logs reduction on concrete. A shortened application procedure with pH‐adjusted bleach resulted in lower efficacy on wood, and a greater apparent potential for contamination spread. Conclusions: Consideration of material surface type is important when selecting a decontaminant. Also, achieving conditions that effectively inactivate surface biological contamination are critical to preventing the spread of contamination. Significance and Impact of the Study: Results presented here are intended to help development of remediation plans following a biological contamination incident.     

Adaptation and cross-adaptation of <Emphasis Type="Italic">Listeria monocytogenes</Emphasis> and <Emphasis Type="Italic">Salmonella enterica</Emphasis> to poultry decontaminants

Information on the potential for acquired reduced susceptibility of bacteria to poultry decontaminants occurring is lacking. Minimal Inhibitory Concentrations (MICs) were established for assessing the initial susceptibility and the adaptative and cross-adaptative responses of four bacterial strains (Listeria monocytogenes serovar l/2a, L. monocytogenes serovar 4b, Salmonella enterica serotype Typhimurium, and S. enterica serotype Enteritidis) to four poultry decontaminants (trisodium phosphate, acidified sodium chlorite -ASC-, citric acid, and peroxyacetic acid). The initial susceptibility was observed to differ among species (all decontaminants) and between Salmonella strains (ASC). These inter- and intra-specific variations highlight (1) the need for strict monitoring of decontaminant concentrations to inactivate all target pathogens of concern, and (2) the importance of selecting adequate test strains in decontamination studies. MICs of ASC (0.17±0.02 to 0.21±0.02 mg/ml) were higher than the U.S. authorized concentration when applied as a pre-chiller or chiller solution (0.05 to 0.15 mg/ml). Progressively increasing decontaminant concentrations resulted in reduced susceptibility of strains. The highest increase in MIC was 1.88 to 2.71-fold (ASC). All decontaminants were shown to cause cross-adaptation of strains between both related and unrelated compounds, the highest increase in MIC being 1.82-fold (ASC). Our results suggest that the in-use concentrations of ASC could, in certain conditions, be ineffective against Listeria and Salmonella strains. The adaptative and cross-adaptative responses of strains tested to poultry decontaminants are of minor concern. However, the observations being presented here are based on in vitro studies, and further research into practical applications are needed in order to confirm these findings.     

Maintenance of Helicobacter pylori cultures in agar stabs

Background: Helicobacter pylori requires frequent passage at 37 °C with reduced oxygen tension to maintain viability, and recovery from frozen stocks can be unpredictable and slow. Agar stab cultures were assessed as a possible means of maintaining viability without the need to passage every 4–7 days. Materials and Methods: Agar stabs prepared from either Brucella or Brain Heart Infusion media were inoculated deeply with H. pylori strains or H. felis and grown under varying conditions for up to 13 weeks. Subcultures were prepared from these stabs at various intervals to test for viability. Results: Established cultures in agar stabs failed to survive at room temperature but did survive at 37 °C with 10% CO₂ for up to 56 days. H. felis remained viable for up to 28 days. No difference was observed between the two media formulations. Conclusion: H. pylori grown in agar stabs remains viable for prolonged periods of time without the need to subculture and may represent an improved method for storing H. pylori for infrequent use.     

Decontamination of a hard surface contaminated with Bacillus anthracisΔSterne and B. anthracis Ames spores using electrochemically generated liquid-phase chlorine dioxide (eClO2)

Aims: To evaluate the inactivation of Bacillus anthracisΔSterne and Ames spores using electrochemically generated liquid‐phase chlorine dioxide (eClO₂) and compare two sporulation and decontamination methods with regard to cost, safety and technical constraints. Methods and Results: Spores were prepared via agar and broth methods and subsequently inoculated and dried onto clean, autoclave‐sterilized glass coupons. Bacillus anthracis spore inactivation efficacy was evaluated using the modified three‐step method (AOAC 2008.05) and a single‐tube extraction method. Spores (7·0 ± 0·5 logs) were inactivated within 1 min at room temperature using freshly prepared eClO₂. Bacillus anthracisΔSterne spores decreased in size after eClO₂ treatment as measured using a Beckman Coulter Multisizer. Conclusions: eClO₂ saturation of a hard surface was an effective B. anthracis sporicide. Broth sporulation and the single‐tube extraction method required less time and fewer steps, yielded a higher percentage of phase‐bright spores and showed higher spore recovery efficiency compared with AOAC 2008.05, making it more amenable to biosafety level 3 (BSL3) testing of virulent spores. Significance and Impact of the Study: Two test methods demonstrated the sporicidal efficacy of eClO₂. A new single‐tube extraction test protocol for decontaminants was introduced.     

Cadmium decontamination and reversal potential of teratological forms of the diatom Planothidium frequentissimum (Bacillariophyceae) after experimental contamination

While the induction of teratology by cadmium (Cd) on diatoms is already known, reversal kinetics are not well documented. This study aims to understand the viability of diatoms exhibiting teratological frustules and their reproduction capacities within a Cd‐impacted population to predict their return to normal diatom forms. We worked on a frequently encountered species in French hydrosystems: Planothidium frequentissimum (Lange‐Bertalot) Round & L. Bukhtiyarova. First, a 21‐d contamination phase highlighted increasing inductionof different teratological types in response to two levels of Cd contamination: 20 and 100 μg · L^?1. The deformity counting indicated that Cd firstly generated striae and mixed teratologies, then affected the central area and the valves. Second, a 28‐d decontamination phase demonstrated the Cd depuration capacity of Planothidium frequentissimum. Cd half‐lives appeared relatively low, ~6 d for the 100 μg · L^?1 condition. Moreover, the decontamination phase showed a decrease in teratology abundances, but a still incomplete recovery after 28 d. Deformations of the striae appeared to be the most sustainable phenotype since they were still significantly higher than in reference cultures at the end of the decontamination phase for both Cd cultures.     

Environmental persistence of Brucella abortus in the Greater Yellowstone Area

Bison (Bison bison) and elk (Cervus elaphus) of the Greater Yellowstone Area (GYA) are the last remaining reservoirs of bovine brucellosis (Brucella abortus) in the United States. An important factor in evaluating the risk of transmission to cattle is the persistence of bacteria and infectious birth materials shed on pastures where cattle graze. We selected 2 study areas near the northern and western boundaries of Yellowstone National Park (YNP) to determine the persistence of bacteria on fetal tissue, soil, and vegetation, and scavenging on infectious materials from birth and abortion sites. We performed 3 independent field experiments to determine: 1) persistence of Brucella abortus (RB51) purposely applied to fetal tissues, 2) scavenging of fetuses by native scavengers, and 3) natural contamination of birth or abortion sites in the GYA. Results from these field experiments established that Brucella bacteria can persist on fetal tissues and soil or vegetation for 21–81 days depending on month, temperature, and exposure to sunlight. Bacteria purposely applied to fetal tissues persisted longer in February than May and did not survive on tissues beyond 10 June regardless of when they were set out. Brucella abortus field strain persisted up to 43 days on soil and vegetation at naturally contaminated bison birth or abortion sites. Fetuses were scavenged by a variety of birds and mammals in areas near YNP and more rapidly inside YNP than outside the Park boundary. Models derived from our data determined a 0.05% chance of bacterial survival beyond 26 days (95% Credible Interval of 18–30 days) for a contamination event in May. May 15 is the final date for hazing all bison into Yellowstone National Park under the current interagency bison management plan. With these data managers can predict when it is safe to graze cattle onto pastures previously occupied by bison. © 2011 The Wildlife Society.     

Effects of acclimation and diapause on the cold tolerance of Trogoderma granarium

Khapra beetle, Trogoderma granarium Everts (Coleoptera: Dermestidae), is a pest of stored grain in Africa, Asia, and Europe. It is a quarantine insect for much of the rest of the world. Control of T. granarium can be achieved with methyl bromide, but this fumigant is an ozone‐depleting substance and is being phased out worldwide. Thus, there is an urgent need to find new methods of control, including the use of low temperatures. Here, we assess the effects of diapause and cold acclimation on the cold tolerance of T. granarium. The percentage of larvae in diapause increased with larval density, reaching 57.3% when reared at a density of 73 larvae g^?1 diet. The cold tolerance of T. granarium was assessed by the supercooling points (SCPs) of various life stages. The SCP of non‐acclimated insects ranged from ?26.2 ± 0.2 °C (mean ± SEM) for eggs to ?14.4 ± 0.4 °C for larvae. The lowest SCP for larvae, ?24.3 ± 0.3 °C, was obtained for diapausing‐acclimated larvae. Based on mean LT₅₀ values, the most cold‐tolerant stage at ?10 °C was the diapausing‐acclimated larvae (87 days) followed by non‐diapausing‐acclimated larvae (51 days), diapausing non‐acclimated larvae (19 days), adults (4 days), non‐diapausing non‐acclimated larvae (2 days), pupae (0.4 days), and eggs (0.2 days). The estimated times to obtain 99.9968% mortality (Probit 9) for diapausing‐acclimated larvae are 999, 442, 347, 84, and 15 days at 0, ?5, ?10, ?15, and ?20 °C, respectively. Probit 9 is an estimated value used by quarantine experts to estimate conditions that are required to kill all insects. In light of the long exposure time needed to control T. granarium even at ?20 °C, cooling to below ?27 °C (i.e., below the SCP of eggs) will quickly kill all life stages and may be the best way to control this insect with low temperatures.     

Life table parameters of the tomato leaf miner Tuta absoluta (Lepidoptera: Gelechiidae) on different tomato cultivars

The tomato leaf miner Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is a devastating pest of tomato plants originating from South America. In this study, the effect of three tomato cultivars on the life table parameters of T. absoluta was investigated. Data were analysed based on age‐stage, two‐sex life table theory. The net reproduction rate (R₀) of T. absoluta on Falkato, Isabella and Grandella cultivars was determined as 15.73 ± 3.35, 17.66 ± 3.33 and 10.03 ± 4.10 eggs, respectively. The intrinsic rate of increase (r) value on those cultivars was 0.09 ± 0.007, 0.095 ± 0.006 and 0.074 ± 0.017 day^?1, respectively. Finite population rates of increase (λ) were 1.095 ± 0.008, 1.099 ± 0.007 and 1.076 ± 0.018 day^?1, and mean generation times (T) on the three cultivars were 30.09 ± 0.29, 30.22 ± 0.27 and 31.12 ± 0.65 days, respectively. The egg incubation period, pre‐adult duration and fecundity on these tomato cultivars was also calculated. The results showed no significant differences between r, R₀, and T on the three tested tomato cultivars. These findings can be useful for developing an integrated pest management strategy against this noxious pest.     

Comparison of culture and PCR for the detection of Brucella melitensis in blood and lymphoid tissues of serologically positive and negative slaughtered sheep

Aims: To compare the culture and PCR methods for detection of Brucella melitensis in blood and lymphoid tissue samples obtained from slaughtered sheep (n = 162) testing positive/negative in serological tests (Rose Bengal test and serum agglutination test). Methods and Results: Of 162 sheep examined, 45 were positive and 117 negative in serological tests. A PCR assay based on a pair of Br. melitensis‐specific primers was used to detect DNA in blood and lymphoid tissue. Brucella melitensis was isolated from 1·2% (2/162) and 17·2% (28/162) of the blood and lymphoid tissue samples respectively. Positive PCR products with a molecular size of 731 bp were obtained from 27·7% (45/162) of blood and 29·0% (47/162) of lymphoid tissue samples. Conclusions: The species‐specific PCR assay detected a higher number of Br. melitensis DNA both from serologically positive (P < 0·01 in blood PCR, P < 0·001 in tissue PCR) and serologically negative (P < 0·001 in both blood PCR and tissue PCR) sheep compared with classical bacteriological culture methods. Significance and Impact of the Study: The results emphasize the importance of using more than one type of diagnostic technique for the detection of animals positive for brucellosis, especially with epidemiological purposes.     

Type III Secretion Homologs Are Present in <Emphasis Type="Italic">Brucella melitensis</Emphasis>, <Emphasis Type="Italic">B. ovis</Emphasis>, and <Emphasis Type="Italic">B. suis</Emphasis> biovars 1, 2, and 3

Protein sequences from characterized type III secretion (TTS) systems were used as probes in silico to identify several TTS gene homologs in the genome sequence of Brucella suis biovar 1 strain 1330. Four of the genes, named flhB, fliP, fliR, and fliF on the basis of greatest homologies to known flagellar apparatus proteins, were targeted in PCR and hybridization assays to determine their distribution among other Brucella nomen species and biovars. The results indicated that flhB, fliP, fliR and fliF are present in Brucella melitensis, Brucella ovis, and Brucella suis biovars 1, 2 and 3. Similar homologos have been reported previously in Brucella abortus. Using RT-PCR assays, we were unable to detect any expression of these genes. It is not yet known whether the genes are the cryptic remnants of a flagellar system or are actively involved in a process contributing to pathogenicity or previously undetected motility, but they are distributed widely in Brucella and merit further study to determine their role. Received: 11 February 2002 / Accepted: 13 June 2002     

Characterization of hematological parameters and blood cells of cultured Gymnocypris eckloni Herzenstein, 1891

The objective of the study was to obtain baseline data on haematological parameters, blood cell sizes and morphology in cultured male and female Gymnocypris eckloni Herzenstein, 1891. Forty‐eight healthy 3‐year‐old G. eckloni (26 males: 525.79 ± 48.56 g weight, 34.51 ± 1.88 cm total length; 22 females: 507.60 ± 54.48 g weight, 33.97 ± 1.84 cm total length) were used for this study. Both male and female gonadal maturity were at stage III (maturing). The fish were reared in 25–36 m² outdoor tanks at dissolved oxygen 6.86 ± 0.48 mg L^?1, pH 7.22 ± 0.58, temperature 12.40 ± 0.94°C and stocking density 50–80 fish m^?3 during November 2014. The fish were fed commercial carp floating foods containing 35% crude protein three times daily. Haematological values were performed manually on heparin anticoagulated blood specimens using standard methods. The morphological features of blood cells and differential cell counts were done on Wright–Giemsa stained blood smears with no anticoagulants. Erythrocytes, leucocytes (neutrophils, eosinophils, lymphocytes and monocytes) and thrombocytes were distinguished and characterized under light microscope. The percentage of the different leukocytes revealed predominance of small lymphocytes (male: 62.31 ± 2.06%; female: 63.00 ± 2.25%) and nurophiles (male: 23.85 ± 1.51%; female: 23.49 ± 1.67%) followed by fewer monocytes (male: 4.81 ± 0.68%; female: 4.80 ± 0.77%) and few eosinophils (male: 3.73 ± 0.82%; female: 3.52 ± 0.67%). The nurophile percentages of each stage showed that metamyelocyte accounted for the most (male: 13.29 ± 0.88%; female: 13.07 ± 0.98%), followed by banded ones (male: 7.18 ± 0.49%; female: 7.00 ± 0.58%). The microstructure of G. eckloni blood cells was similar to that of other fish. Sex‐dependent differences for the erythrocyte counts, haemoglobin, haematocrit and mean corpuscular haemoglobin were found (P < 0.05 or P < 0.01); while differences in other haematological parameters (P > 0.05) and blood cell morphology between male and female fish were not significant. Hematologic parameters and knowledge of morphological characteristics of male and female G. eckloni blood cells could be utilized to evaluate the health status of this species in captivity.     

Survival and oviposition of a western corn rootworm variant feeding on soybean

The role of soybean, Glycine max (L.) Merril (Fabaceae), in the circumvention of crop rotation was evaluated by observing the effects of soybean herbivory on western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) survival and oviposition. Field collected D. virgifera adults resistant to annual rotation of corn, Zea mays (L.) (Poaceae), and soybean were given the opportunity to feed on a soybean pre‐treatment diet prior to diet combinations of corn, soybean, and/or water. The number of days that the adults survived after removal of the diet combinations was used as a measure of insect vigor and diet quality. Willingness to feed on a soybean foliage pre‐treatment did not indicate greater ability to benefit nutritionally from soybean herbivory. Adult D. virgifera previously feeding on corn, soybean, or water survived starvation for 8.3 ± 0.15 days, 5.1 ± 0.13 days, and 3.9 ± 0.14 days, respectively. Diabrotica virgifera recover from 2 days of starvation or poor diet if subsequently given access to corn tissues for 2 days. Eggs were laid within 1 day of access to poor diet or starvation. Other D. virgifera, captured as they flew from a cornfield into a soybean field, were maintained on soybean foliage or only water until they died. Of the beetles with access to soybean foliage, 24% fed within 24 h after capture and survived 1 day longer than insects given only water. Only 20% of these D. virgifera were able to lay eggs without consuming additional corn prior to death. Few D. virgifera enter soybean fields capable of immediate oviposition. Female reproductive status and diet quality influence the likelihood of oviposition following exposure to stress. The effects of soybean herbivory may contribute to the proximate mechanism of resistance to crop rotation.     

Effects of desiccation and rewetting on the release and decomposition of dissolved organic carbon from benthic macroalgae

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Optimizing acidified bleach solutions to improve sporicidal efficacy on building materials

Aims: We evaluated whether lowering pH (with acetic acid) and raising free available chlorine (FAC) levels in bleach solutions would improve efficacy in inactivating Bacillus spores on different materials. We also determined how varying pH and FAC levels affected bleach stability. Methods and Results: Acidified bleach solutions with pH levels of 4·5, 6 and 7·5 and FAC levels between 5000 and 10 000 ppm were evaluated for decontamination efficacy against Bacillus subtilis spores inoculated onto test coupons made from wood, ceramic and galvanized steel. Lowering the pH or increasing the FAC level improved efficacy in some of the tests, but depended on the material, which significantly affected decontamination efficacy. The acidified bleach at pH of 7·5 was significantly less effective than bleach at a pH of 4·5 or 6. The FAC levels in the bleach were the most stable at pH 4·5, and stability at pH 4·5 was not significantly affected by the initial FAC level. Conclusions: It may be advisable to use bleach solutions with lower pH (rather than high FAC levels) in light of both the decontamination efficacy and bleach stability results. For wood materials, use of sporicides other than acidified bleach may be warranted. Significance and Impact of the Study: These results may be useful in preparing acidified bleach solutions for decontamination of materials contaminated with spores such as Bacillus anthracis.     

Comparison of decontamination methods for primary isolation of Mycobacterium bovis in paucibacillary bovine tissues

Aims: To compare three decontamination methods applied to paucibacillary samples for primary isolation of Mycobacterium bovis from suspect lesions. Tuberculosis caused by Myco. bovis is an important infectious disease of cattle in Brazil and also has zoonotic potential. Although a national campaign based on testing and slaughtering cattle has achieved good results, there is a strong need to develop better diagnostic methods to identify cattle with recent infections harbouring few bacilli. Methods and Results: A dairy herd (274 adult crossbred cows) located in the state of Rio de Janeiro was tested for tuberculosis with both single intradermal tuberculin test and comparative intradermal tuberculin test. Reactive cows (n = 27, 9·8%) were slaughtered and suspect lesions were collected (one sample per cow). Samples considered paucibacillary (based on microscopy) were decontaminated with 0·75% hexadecylpyridinium chloride (HPC), 4% sodium hydroxide (Petroff) or 6% sulphuric acid. Using these methods, 10, five and six, respectively, of the 27 samples yielded positive cultures. Overall, Myco. bovis was isolated from 14 of 24 cows. Although the HPC method resulted in isolation of more Myco. bovis strains than either Petroff or sulphuric acid methods (P = 0·015), it did not result in the recovery of Myco. bovis from all samples. However, using both HPC and 6% sulphuric acid methods for decontamination was possible to identify 13 of 14 (92·9%) of infected cows. Conclusions: At least two methods should be used concurrently for primary isolation of Myco. bovis from bovine tissues, particularly for paucibacillary samples. Significance and Impact of the Study: Detection of low numbers of Myco. bovis in tissue is an important goal in optimizing the detection of bovine tuberculosis and should assist in identification of infected cattle, in particular, those with few Myco. bovis bacilli. This was apparently the first study comparing three decontamination methods for the detection of Myco. bovis in paucibacillary samples from naturally infected cattle.     

Participation of the molecular chaperone DnaK in intracellular growth of Brucella suis within U937-derived phagocytes

In the intracellular bacterium Brucella suis, the molecular chaperone DnaK was induced under heat-shock conditions and at low pH. Insertional inactivation of dnaK and dnaJ within the dnaK/J locus led to the conclusion that DnaK, but not DnaJ, was required for growth at 37°C in vitro. Viability of the dnaK null mutant was also greatly affected at low pH. Under conditions allowing intracellular multiplication, the infection of U937-derived phagocytes resulted in long-lasting DnaK induction in the wild-type bacteria. In infection experiments performed with both mutants at the reduced temperature of 30°C, the dnaK mutant of B. suis survived but failed to multiply within U937 cells, whereas the wild-type strain and the dnaJ mutant multiplied normally. Complementation of the dnaK mutant with the cloned dnaK gene restored growth at 37°C, increased resistance to acid pH, and increased intracellular multiplication. This is the first report of the effects of dnaK inactivation in a pathogenic species, and of the temperature-independent contribution of DnaK to intracellular multiplication of the pathogen B. suis.     

Ecological and morphological characteristics of the endoparasitoids of larval Acronicta rumicis (Lepidoptera: Noctuidae)

The Hymenopterans Glyptapanteles liparidis, Microplitis sp. and Diadegma sp. were found to be larval parasitoids and koinobionts of Acronicta rumicis (Lepidoptera: Noctuidae). Mesochorus semirufus is believed to be a new unreported hyperparasitoid of G. liparidis, which, along with M. semirufus, is a gregarious parasitoid. In contrast, the parasitoids Microplitis sp. and Diadegma sp. are solitary. All of the hymenopteran parasitoids are multivoltine insects that emerge from A. rumicis more than once. Compcilura concinnata, Euexorista sp. and Exorista sp. of the Diptera were found to be larval–pupal parasitoids, solitary parasitoids and koinobionts. These three species are univoltine, and emerge only once from A. rumicis. Morphological and life cycle data were collected for G. liparidis, and for the parasitoids of that species found in this study. The major and minor axes of an egg of G. liparidis were 0.10 and 0.02 mm, respectively, while the mean clutch size of G. liparidis was 67.71 ± 39.36 individuals. The body length of female and male G. liparidis were 2.25 ± 0.06 and 2.21 ± 0.12 mm, respectively, and the longevity of an adult was 2.93 ± 0.96 days. Among the parasitoids, the mean body length of an adult Microplitis sp. was 3.5 mm and adults lived for an average of 8.13 ± 3.54 days. The adult Diadegma sp. was larger (mean body length 6.5 mm) but lived for a shorter interval (3.33 ± 1.32 days). The body lengths of female and male M. semirufus were 3.16 ± 0.11 and 3.10 ± 0.23 mm, respectively, greater than the body lengths of female and male G. liparidis. The body lengths of adult C. concinnata, Euexorista sp. and Exorista sp. were 9.5, 9.53 and 8.68 mm, respectively. All of their pupae were dark brown.     

Anesthesia of wild female Persian sturgeon,Acipenser persicus (Borodin, 1897) breeders during controlled propagation: effects on hematological parameters,stress response and reproductive performance

This study examined the effects of anesthesia on the hematological and biochemical parameters as well as the reproductive performance of wild female Persian sturgeon, Acipenser persicus, during controlled spawning. Fourteen mature females were divided into two groups: ‘anesthetized’ and ‘non‐anesthetized’. All activities including transportation, catheterizing and handling were performed with both groups: (i) under anesthesia (150 ppm clove oil), and (ii) without anesthesia. After 10 days storage and handling, blood samples were taken from all fish after anesthesia. No significant differences were found in the reproductive performance of either group. However, differences were found in the hematological parameters, with values being significantly higher in the non‐anesthetized group, including neutrophils (34.36 ± 6.33% vs 23.63 ± 5.22%), monocytes (2.84 ± 1.70% vs 1.27 ± 0.64%), mean corpuscular volume (321.3 ± 39.40 pg vs 228.0 ± 24.46 pg) and mean corpuscular hemoglobin (106.9 ± 15.70 fl vs 76.50 ± 7.50 fl). Significantly lower values were found in the non‐anesthetized group for lymphocytes (60.68 ± 7.25% vs 73.54 ± 4.80%), Hb (4.62 ± 0.74 mg dl^?1 vs 6.28 ± 1.21 mg dl^?1), Hct (13.86 ± 1.76% vs 18.84 ± 3.85%), red blood cell (0.43 ± 0.05 cell mm^?3 vs 0.85 ± 0.13 × 10⁶ cell mm^?3) and white blood cell (22 403 ± 2240 cell mm^?3 vs 35 318 ± 3084 cell mm^?3). The non‐anesthetized fish had significantly higher cortisol levels compared to the anesthetized group (62.33 ± 8.85 ng ml^?1 vs 46.12 ± 8.07 ng ml^?1). There was no difference in plasma glucose levels between groups. It is concluded that the use of clove oil as an anesthetic is suitable for handling of wild female Persian sturgeon in controlled propagation programmes. However, further research is needed to determine a standardized protocol for the safe application of anesthesia for use in sturgeons in general.