The diet of the parasitic mite Trichosurolaelaps crassipes Womersley, 1956 and its potential to act as a disease vector

The anatomy of the gaster of Trichosurolaelaps crassipes is illustrated. Protonymphs, deutonymphs and adults were all seen with a scarlet gut and gave strongly positive results to occult blood tests. Host blood cells were identified in a gut smear from a female mite. An adult female gut could hold about 7.1 x 10(-6) ml. The mite was seen frequenting small open skin lesions. Gram-negative bacteria identified in the mite's gut are most likely to be endosymbionts. Mite larvae have their gastric caeca invaded by these bacteria before birth. There is a potential for this mite to act as a weak vector of disease from one possum to another.     

Communications: Bacteria in the Hemolymph of the Freshwater Prawn Macrobrachium rosenbergii

Abstract

Bacteria from the hemolymph of the freshwater prawn Macrobrachium rosenbergii were identified and quantified. Total bacterial counts ranged from 0.0 to 4.6 × 10⁵ cells/1.0 mL of hemolymph. Predominant bacteria isolated included Aeromonas spp., Bacillus sp., and Pseudomonas spp. No bacteria were found in the hemolymph of prawns without lesions. The predominant species of bacteria isolated from water samples of prawn culture ponds was a chitinoclastic Bacillus sp.     

肺炎克雷伯氏菌强毒株的分离鉴定及16-23SrRNAITS序列分析

为确诊疑似仔猪肺炎克雷伯氏菌(K.pneumonia)感染,并研究其病原的致病性、耐药性、16-23SrRNA ITS系统进化特征,本研究从云南因肺炎、腹泻而大量死亡的仔猪中分离到1株革兰氏阴性短粗杆菌,命名为KP14013,对其进行生化鉴定、16SrRNA鉴定,研究其对小白鼠和仔猪的致病性,并对其16-23SrRNA ITS基因进行测序和遗传进化分析。结果显示,KP14013分离株生化特征与肺炎克雷伯氏菌相符,其16SrRNA与GenBank中23株肺炎克雷伯氏菌代表株之间的同源性均为99%,将KP14013鉴定为肺炎克雷伯氏菌。KP14013对小白鼠半数致死量(LD50)为3×101.8 CFU,腹腔注射3×108 CFU可使仔猪100%致死。16-23SrRNA ITS系统进化关系结果表明,KP14013与GenBank中收录的15株肺炎克雷伯氏菌形成进化树的一个分支,属于同一个亚群,它们之间的核苷酸同源性为98.4%~99.2%。本研究证实了肺炎克雷伯氏菌是该起仔猪腹泻大量死亡的病原;KP14013分离株为毒力极强菌株,具有多重耐药性,其16-23SrRNA ITS与GenBank中收录的肺炎克雷伯氏菌代表株之间核苷酸存在差异,可用于肺炎克雷伯氏菌菌株间的鉴别。     

House dust and storage mite contamination of dry dog food stored in open bags and sealed boxes in 10 domestic households

Dry pet food is a potential source of exposure to house dust and storage mite allergens in canine atopic dermatitis. This study evaluated contamination of house dust and dry dog food stored in paper bags, sealable plastic bags and sealable plastic boxes in 10 households for 90 days using Acarex^® tests for guanine, a Der p 1 ELISA and mite flotation. Acarex^® tests were negative in all the food samples but positive in all the house dust samples. The Der p 1 levels and mite numbers significantly increased in food from paper bags (P = 0.0073 and P = 0.02, respectively), but not plastic bags or boxes. Mite numbers and Der p 1 levels were 10–1000 times higher in house dust than the corresponding food samples (P < 0.0001). There were significant correlations between Der p 1 in house dust and food from the paper (P < 0.0001) and plastic bags (P = 0.003), and mite numbers in house dust and food from the paper bags (P = 0.0007). Bedding and carpets were significantly associated with Der p 1 levels in house dust (P = 0.015 and P = 0.01, respectively), and food from the paper (both P = 0.02) and plastic bags (P = 0.03 and P = 0.04, respectively). Mites were identified in six of 10 paper bag, three of 10 plastic bag, one of 10 plastic box and nine of 10 house dust samples. These comprised Dermatophagoides (54%), Tyrophagus (10%; all from food) and unidentified mites (36%). Storage of food in sealable plastic boxes largely prevented contamination for 3 months. Exposure to mites and mite proteins in all the stored food, however, appeared to be trivial compared with house dust.     

Pathogenesis of Enteric Septicemia of Channel Catfish,Caused by Edwardsiella ictaluri: Bacteriologic and Light and Electron Microscopic Findings

Abstract

To clarify early events in the pathogenesis of enteric septicemia of catfish, 140 channel catfish Ictalurus punctatus (8–10 months old) were each infected with approximately 1.0 × 10⁹ colony-forming units of Edwardsiella ictaluri by intragastric intubation. Fish were sacrificed at 0, 0.25, 0.5, 1, 3, 6, 12, 24, 48, 72, 96, and 120 h postinfection (PI). Multiple tissue samples at all scheduled sampling times were evaluated by gross observation, light and electron microscopy, and immunohistochemical methods. In addition, at each sampling time, stomach, intestine, trunk kidney, and liver were cultured to quantitate bacteria. Trunk kidney cultures were positive by 0.25 h PI, indicating rapid transmucosal passage. In the intestine, E. ictaluri was seen in contact with the brush border at 0.5 h PI. Also at 0.5 h PI, dilated basilar cells with large intracytoplasmic inclusions were observed adjacent to the basement membrane. From 1 to 3 h PI, occasional necrotic enterocytes were seen on tips of intestinal folds. Proprial leukocytes were rare before 24 h PI but common thereafter. Immunoelectron microscopy showed E. ictaluri in vacuoles within phagocytes as early as 24 h PI in the intestinal mucosa. In other tissues, earliest observed microscopic lesions (48 h PI) consisted of bacteria within vacuoles of phagocytic cells contained within blood vessels. Bacteria were also seen within degenerate vacuoles in enterocytes and hepatocytes at 72, 96, and 120 h PI. This study confirms that E. ictaluri can invade channel catfish within 0.25 h PI by crossing the intestinal mucosa and suggests that the bacterium may have invasion and survival strategies similar to those of other enteroinvasive members of the Enterobacteriaceae.     

Identification of lactic acid bacteria in the feces of dairy cows fed whole crop maize silage to assess the survival of silage bacteria in the gut

In order to assess the survival of lactic acid bacteria (LAB) in whole crop maize silage in the gut of dairy cows, one representative silage sample and three different feces samples were collected from dairy cows on three dairy farms in Hua Bei, China and three dairy farms in Kyushu, Japan. The composition of the bacterial community was examined by denaturing gradient gel electrophoresis and quantitative polymerase chain reaction. Lactobacillus acetotolerans was detected in all bunker‐made maize silage samples, regardless of the dairy farm or sampling region from which they were sourced. A total of eight LAB species were detected in the maize silage samples, of which three (L. acetotolerans, L. pontis and L. casei) appeared to survive digestion. The populations of L. acetotolerans in silage and feces were 10^6–7 and 10^3–4 copies/g, respectively, indicating that, even for the LAB species showing potential survival in the gut, competition in this niche may be harsh and the population may substantially decrease during the digestion process. It may be difficult for silage LAB to survive in the gut of silage‐fed dairy cows, because marked decrease in population can take place during the digestion process, even for surviving species.     

Active components of essential oils as acaricides against Dermanyssus gallinae

ABSTRACT

1. The aim was to evaluate the acaricidal effects of pure active components of essential oils against poultry red mite (Dermanyssus gallinae) as an alternative to chemical acaricides (organophosphates and pyrethroids).

2. The toxicities of five pure active components of essential oils (eugenol from clove bud, eucalyptol from rosemary, limonene from citrus fruits, linalool from lavender and cinnamaldehyde from cinnamon) were tested on D. gallinae females in an impregnated paper assay.

3. The active substances were dissolved in water and Tween 20 and applied at concentrations ranging from 0.002 to 0.06 µl/cm². Toxicity was expressed as a lethal dose (LD₅₀ or LD₉₀).

4. The highest mortality was observed with eugenol. The LD₉₀ was estimated to be 5.1 µg/cm² for this substance, followed by cinnamaldehyde, the LD₉₀ of which was estimated to be 11.0 µg/cm². Limonene and eucalyptol were generally less effective in controlling D. gallinae.     

<Emphasis Type="Italic">Streptococcus uberis</Emphasis>-specific T cells are present in mammary gland secretions of cows and can be activated to kill <Emphasis Type="Italic">S. uberis</Emphasis>

The presence, phenotype and function of Streptococcus uberis-specific T cells in the mammary gland secretion (MGS) and blood of cows exposed to S. uberis were assessed. MGS T cells in the udder were purified and incubated with autologous blood monocytes as antigen-presenting cells (APC). Most cows, irrespective of prior S. uberis infection status and lactation status, were shown to have S. uberis-specific T cells both in MGS and in the blood. When cells from a subgroup of cows were studied, it was found that the S. uberis-specific T cells produced high levels of interferon-gamma (IFN-γ), but low levels of interleukin-10 (IL-10). A high percentage of responding T cells were of the CD8 ⁺ memory (CD45RO) subset. T cells from the MGS specific for S. uberis were propagated from animals during the drying off period and expanded in vitro using interleukin-2 (IL-2) and S. uberis antigens. This led to the accumulation of T cells of the CD8 ⁺ subset bearing memory cell markers (CD45A ⁻ , CD45RO ⁺ ), which released high levels of IFN-γ. Four of the five T cell lines derived from the MGS of three animals had substantial direct killing activity towards S. uberis in vitro. It is concluded that there is an emergence of S. uberis-specific bactericidal T cells in the MGS of cows after infection or environmental exposure to S. uberis. Vaccines aimed at activating and expanding this T cell population in the mammary glands of cattle may offer an avenue for the prevention of mastitis caused by S. uberis.     

Development of livestock feed additives using porcine blood cells

Livestock blood discarded during slaughtering has potentially valuable components such as plasma proteins and haemoglobin. Plasma is used as a feed additive following processing via different methods, including spray drying, whereas blood cells have been underutilized. In this study, we developed haemoglobin hydrolysate (HH) and iron-enriched residue (IER) from porcine blood cells and investigated whether their oral administration regulates the immune system and gut microbiota in growing rats. Twenty-one Sprague-Dawley male rats (n = 7) were used during a 4-week trial and were fed a control, HH or IER diet. The ratio of beneficial bacteria such as Lactobacillus and Akkermansia strains increased in rats fed HH or IER diets. Moreover, compared with the control group, the IER group had an elevated ratio of Lactobacillus to Enterobacteria, which is regarded as an index of beneficial aspect in the gut. Phagocytosis of peripheral blood leucocytes was higher in the HH and IER groups than in the control group. The level of plasma immunoglobulin G increased to approximately 72.7 mg/ml and 152.0 mg/ml in the HH and IER groups, respectively, which was significantly (p < 0.05) higher than that in the control group. These results confirm that HH and IER developed in this study may be a potential additive for animal feeds.     

Microflora in traditional starter cultures for fermented milk, hurunge, from Inner Mongolia, China

Microflora were investigated in traditional starter cultures for fermented milk, hurunge, which are used for fermented dairy products by nomadic families in the Inner Mongolia Autonomic Region, China. The acid‐forming bacteria and yeast counts ranged from 1.8 × 10⁵ to 5.3 × 10⁸ c.f.u./g and from 6.1 × 10⁵ to 3.2 × 10⁶ c.f.u./g, respectively. Sixty‐six strains of lactic acid bacteria and 30 strains of yeasts were isolated and identified from three hurunge samples collected from the nomadic families. Lactococcus raffinolactis was the most predominant lactococci isolated from these samples. The other lactococci were Lactococcus lactis ssp. lactis, Lactococcus lactis ssp. cremoris, and Leuconostoc mesenteroides ssp. cremoris. Two major lactobacilli strains, Lactobacillus plantarum and Lactobacillus casei, were identified. In addition, Lactobacillus kefiranofaciens, Lactobacillus acetotolerans, which grew in 11% acetic acid culture medium, and Lactobacillus homohiochii, which grew in the culture medium containing 16% ethanol, were also identified. The isolated yeast strains were identified as Candida kefyr, Saccharomyces cerevisiae, Kluyveromyces marxianus var. lactis, Candida krusei and Candida valida.     

The epidermal location and possible feeding site of Psorergates ovis, the sheep itch mite

Skin biopsies from two Merino sheep heavily infested with Psorergates ovis were immersed in liquid nitrogen and cut into vertical frozen sections stained with lipophilic Sudan IV or Oil Red O and haematoxylin. A survey of the lateral distribution of mite sections showed a majority (ca 80%) were in or within 0.2 mm of the follicle mouth. A survey of vertical distribution showed no mite penetration deeper than inner stratum corneum where 57% of mite sections were seen; 30% were within outer stratum corneum or scurf; 13% were on the outer surface and less than 1% were detached. Lipid was the only material seen within stained mites at a location considered to be gut. This was supported by dosing sheep with quinacrine, taking biopsies at Day 6 and Day 14 and examining frozen sections under blue light. Fluorescent lipid was seen at a location considered to be mite gut. From these results it was recommended that acaricide treatments against P. ovis be lipophilic and administered transepidermally because less than 15% of mites were at a superficial location likely to be reached by topical application.     

Diversity of bacterial pathogens and their antimicrobial resistance profile among commensal rodents in Qatar

Rodents are sources of many zoonotic pathogens that are of public health concern. This study investigated bacterial pathogens and assessed their antimicrobial resistance (AMR) patterns in commensal rodents in Qatar. A total of 148 rodents were captured between August 2019 and February 2020, and blood, ectoparasites, and visceral samples were collected. Gram-negative bacteria were isolated from the intestines, and blood plasma samples were used to detect antibodies against Brucella spp., Chlamydophila abortus, and Coxiella burnetii. PCR assays were performed to detect C. burnetii, Leptospira spp., Rickettsia spp., and Yersinia pestis in rodent tissues and ectoparasite samples. Antimicrobial resistance by the isolated intestinal bacteria was performed using an automated VITEK analyzer. A total of 13 bacterial species were isolated from the intestine samples, namely Acinetobacter baumannii, Aeromonas salmonicida, Citrobacter freundii, Citrobacter koseri, Enterobacter aerogenes, Enterobacter cloacae, Escherichia coli, Hafnia alvei, Klebsiella pneumoniae, Providencia stuartii, Proteus mirabilis, Pseudomonas aeruginosa, and Salmonella enterica. The majority of them were E. coli (54.63%), followed by P. mirabilis (17.59%) and K. pneumoniae (8.33%). Most of the pathogens were isolated from rodents obtained from livestock farms (50.46%), followed by agricultural farms (26.61%) and other sources (22.94%). No antibodies (0/148) were detected against Brucella spp., C. abortus, or C. burnetii. In addition, 31.58% (6/19) of the flea pools and one (1/1) mite pool was positive for Rickettsia spp., and no sample was positive for C. burnetii, Leptospira spp., and Y. pestis by PCR. A total of 43 (38%) bacterial isolates were identified as multidrug resistant (MDR), whereas A. salmonicida (n?=?1) did not show resistance to any tested antimicrobials. Over 50% of bacterial MDR isolates were resistant to ampicillin, cefalotin, doxycycline, nitrofurantoin, and tetracycline. The presence of MDR pathogens was not correlated with rodent species or the location of rodent trapping. Seven (11.86%) E. coli and 2 (22.2%) K. pneumoniae were extended-spectrum beta-lactamases (ESBL) producers. These findings suggest that rodents can be a source of opportunistic bacteria for human and animal transmission in Qatar. Further studies are needed for the molecular characterization of the identified bacteria in this study.

     

Presence and diversity of mixed avian Plasmodium spp. infections in introduced birds whose distribution overlapped with threatened New Zealand endemic birds

ABSTRACT

Aims: To determine the presence of infection and co-infection of Plasmodium lineages in introduced birds at translocation sites for the North Island saddleback (Philesturnus rufusater), to investigate their role as Plasmodium spp. reservoirs.

Methods: Blood samples were collected from introduced bird species, with a special focus on blackbirds (Turdus merula) and song thrushes (Turdus philomelos), at six locations in the North Island of New Zealand that were the origin, or translocation sites, for North Island saddleback. Where available, blood smears were examined, and blood samples were tested using nested PCR with subsequent sequence analysis, for the presence of Plasmodium spp.

Results: Of the 55 samples tested using PCR analysis, 39 (71%) were positive for Plasmodium spp., and 28/40 (62%) blood smears were positive for Plasmodium spp. Overall, 31 blood samples were from blackbirds with 28/31 (90%) samples positive for Plasmodium spp. Six distinct avian Plasmodium lineages were identified, including three cosmopolitan lineages; Plasmodium vaughani SYAT05 was detected in 16 samples, Plasmodium matutinum Linn1 in 10 samples and Plasmodium elongatum GRW6 in eight samples. Mixed infections with more than one lineage were detected in 12 samples. Samples from two Australian magpies (Gymnorhina tibicen) were positive for Plasmodium. sp. lineage MYNA02, previously not identified in New Zealand.

Conclusions and clinical relevance: This is the first report from New Zealand in which specific Plasmodium spp. mixed infections have been found in introduced birds. Co-infections with several cosmopolitan Plasmodium lineages were identified, as well as the first report in New Zealand of an exotic avian Plasmodium sp. lineage, in Australian magpies. Whilst the role of introduced birds in maintaining and spreading pathogenic avian malaria in New Zealand is unclear, there is a potential infection risk to native birds, especially where distributions overlap.     

Evaluation of butorphanol, medetomidine and midazolam as a reversible narcotic combination in free-ranging African lions (Panthera leo)

ObjectiveTo evaluate the effects of the combination butorphanol, medetomidine and midazolam (BMM) and its reversibility in lions.Study designProspective clinical trial.AnimalsThirty free-ranging lions, 10 male and 20 female, weighing 81-210 kg.MethodsLions were immobilised with butorphanol mean 0.31 ± SD 0.034 mg kg^?1, medetomidine 0.052 ± 0.006 mg kg^?1, midazolam 0.21 ± 0.024 mg kg^?1 and hyaluronidase 1250 IU administered intramuscularly with a dart gun. Upon recumbency, physiological parameters and anaesthetic depth were monitored 10-15 minutes after darting (T1) and repeated every 10 minutes for a further 30 minutes (T2, T3, T4). Arterial blood gas analyses were performed at T1 and T4. At the end of the procedure, 45-60 minutes after initial darting, immobilisation was reversed with naltrexone 0.68 ± 0.082 mg kg^?1, atipamezole 0.26 ± 0.031 mg kg^?1, and flumazenil 0.0032 ± 0.0007 mg kg^?1 administered intravenously and subcutaneously.ResultsThe BMM combination rapidly induced immobilisation and lateral recumbency was reached within 7.25 ± 2.3 minutes. Median induction score [scored 1 (excellent) to 4 (poor)] was 1.4 (range 1-2). Cardio-respiratory parameters were stable. Heart rate varied from 32 to 72 beats per minute, respiratory rate from 14 to 32 breaths minute^?1 and rectal temperature from 36.6 to 40.3 °C. No sudden arousals were observed. Arterial blood gas analyses revealed a mean pH of 7.33, PaCO₂ of 33 mmHg and PaO₂ of 87 mmHg. Mild to moderate hypoxemia was seen in four lions. Recovery was smooth and lions were walking within 4.4 ± 4.25 minutes. Median recovery score [scored 1 (excellent) to 4 (poor)] was 1.3 (range 1-2).Conclusion and clinical relevanceThe drug combination proved to be effective in immobilising free-ranging healthy lions of both sexes with minimal cardio-respiratory changes.     

Detection of Leishmania (L.) infantum in stray dogs by molecular techniques with sensitive species-specific primers

Background: Canine visceral leishmaniasis (CVL) is a worldwide parasitic zoonosis caused by Leishmania (Leishmania) infantum around the world. Canids are the definitive hosts and sand flies the intermediate hosts.

Objective: To test the hypothesis that a new species-specific primers (Lch14:Lch15, targeting a multiple alignment for L. infantum kDNA minicircle) is an efficient diagnostic tool for L. infantum.

Methods: The presence of L. infantum DNA was assessed in blood samples of 69 stray dogs using the conventional PCR (cPCR) and quantitative PCR (qPCR). Additional 50 lymph nodes and 50 bone marrow samples (positive and negative samples for parasitological tests) from dogs from endemic and nonendemic areas for CVL were also used.

Results: L. infantum strains, and all positive lymph node and bone marrow samples for parasitological test gave positive results for cPCR and qPCR, presenting analytical sensitivity of ～10⁰ parasite mL^?1. For the blood samples, 40/69 (58%; CI 95%; 46%–69%) resulted positive for L. infantum in both tests. All positive samples were confirmed by sequencing.

Conclusion: This study showed the importance of the specific detection of L. infantum based on species-specific primers by molecular techniques, highlighting the application as a confirmation method in epidemiological studies and to adopt the best control measures.     

Neutrophil Phagocytosis Following Inoculation of Salmonella choleraesuis into Swine

Neutrophils are an important mediator of host defence, especially in early stages of infection. A major function of neutrophils is the uptake and killing of invading microbes. Little is known about the effect of neutrophil activity on the pathogenesis and development of the carrier state in swine following infection with Salmonella choleraesuis. A human whole-blood microassay using flow cytometry was modified to measure the effect of S. choleraesuis infection in vivo on the rate of ingestion, or rate of uptake, of homologous bacteria by porcine neutrophils. Pigs were inoculated intranasally with 5–8×10⁸ CFU S. choleraesuis and blood was collected in heparinized tubes at –5, 0, 1, 2, 3 and 4 days post inoculation (PI). Heat-killed S. choleraesuis were labelled with fluorescein isothiocyanate and incubated for various times with diluted whole blood. Red blood cells were lysed, external non-phagocytized bacteria were quenched with a commercially available lysing solution, and fluorescence from internalized bacteria labelled with fluorescein isothiocyanate was detected by flow cytometry. The rate of uptake by neutrophils did not increase until 2 days PI and then remained elevated to 4 days PI. The minimal uptake of S. choleraesuis early after exposure to these organisms may provide an opportunity for the pathogen to colonize and/or replicate to levels that facilitate establishment of a carrier state or clinical infection in swine.     

Biochemical Aspects of Schistosome Behaviour

The haematophagous blood tiematodes that comprise the genus Schistosoma live in a stable environment with an adequate food supply and an efficient waste disposal system.

The main food is glucose obtained from the host blood and assimilated through the tegument Other dietary factors are assimilated through the tegument or the gut epithelium. The male fluke supplements and augments the diet of the female who is thus able to concentrate on egg production.

Live flukes resist attack by the host by secreting an immunologically inert envelope and by incorporating unchanged host serum protein into their structures. They can also incorporate host structural fatty acids unchanged.     

Influences of quorum‐quenching probiotic bacteria on the gut microbial community and immune function in weaning pigs

The aim of this study is to investigate the dynamic gut microbial diversity in weaning swine after administering feed supplemented with probiotic bacteria that specifically inhibit the activity of quorum molecules. Initially, the universal quorum molecule autoinducer‐2 (AI‐2) bioassay results indicated that AI‐2 activity was profoundly inhibited in enterohemorrhagic Escherichia coli (EHEC) O157:H7 in the presence of Lactobacillus acidophilus strain 30SC cell extract, although the growth of EHEC was not affected. Based on plate counting results, bacterial community analysis revealed a specific reduction in coliforms compared to the control, whereas the population of lactobacilli increased in weaning swine in in vivo trials. Supplementation with L. acidophilus strain 30SC did not affect the counts of other communities, such as total aerobes and yeast/mold. In addition, PCR‐denaturing gradient gel electrophoresis analysis showed a significant difference in the 16S rRNA gene products after administering L. acidophilus strain 30SC. Selected bands were sequenced, and most of them were identified as uncultured bacterium clones or a Lactobacillus‐ and Bifidobacterium‐specific community. Therefore, our results indicate that quorum‐quenching probiotic bacteria can significantly modulate the gut microbiota of swine and these beneficial effects can contribute to the improvement of performance and health in the gastrointestinal tract of weaning pigs.