T lymphocyte responses during early enteric Mycobacterium avium subspecies paratuberculosis infection in cattle

Johne's disease (JD) is a costly intestinal disease of ruminants caused by Mycobacterium avium subspecies paratuberculosis (Map), which is transmitted to perinatal calves by the fecal-oral route. Disease control efforts focus on identification and culling of infected cattle from herds; therefore failure to identify animals early is a major obstacle to reducing transmission. Development of host immunity during early JD remain incompletely characterized so detecting subclinical JD using immunologic techniques is a substantial challenge in the field. Development of a test with high sensitivity and specificity is a major research goal with significant implications for the cattle industry. The objectives of this study were to compare early Map-specific T lymphocyte responses in naive, experimentally Map infected and Map vaccinated calves using a subcutaneous matrigel biopolymer-based assay. We examined the phenotype of recruited lymphocytes and local interferon gamma (IFNγ) production within subcutaneously placed matrigel containing Map antigen 30 days after experimentally induced intestinal Map infection or Map vaccination. We show that IFNγ-secreting CD4+ T cells are recruited to matrigel sites in vaccinated but not infected or naïve calves. γδ T cells recruited to matrigel sites of Map-infected calves were mostly WC1-, while γδ T cells recruited to matrigel sites of Map-vaccinated calves were predominantly WC1+. IFNγ at matrigel sites was a discriminating factor between infected calves, naïve calves and vaccinated calves. These data contribute to our understanding of early anti-Map immunity, and may be useful for detecting early intestinal Map infections in calves or for enhancing our ability to discriminate between Map-infected and Map-vaccinated calves.     

Kisspeptin,c‐Fos and CRFR Type 2 Co‐expression in the Hypothalamus after Insulin‐Induced Hypoglycaemia

Normal reproductive function is dependent upon availability of glucose and insulin‐induced hypoglycaemia is a metabolic stressor known to disrupt the ovine oestrous cycle. We have recently shown that IIH has the ability to delay the LH surge of intact ewes. In the present study, we examined brain tissue to determine: (i) which hypothalamic regions are activated with respect to IIH and (ii) the effect of IIH on kisspeptin cell activation and CRFR type 2 immunoreactivity, all of which may be involved in disruptive mechanisms. Follicular phases were synchronized with progesterone vaginal pessaries and at 28 h after progesterone withdrawal (PW), animals received saline (n = 6) or insulin (4 IU/kg; n = 5) and were subsequently killed at 31 h after PW (i.e., 3 h after insulin administration). Peripheral hormone concentrations were evaluated, and hypothalamic sections were immunostained for either kisspeptin and c‐Fos (a marker of neuronal activation) or CRFR type 2. Within 3 h of treatment, cortisol concentrations had increased whereas plasma oestradiol concentrations decreased in peripheral plasma (p < 0.05 for both). In the arcuate nucleus (ARC), insulin‐treated ewes had an increased expression of c‐Fos. Furthermore, the percentage of kisspeptin cells co‐expressing c‐Fos increased in the ARC (from 11 to 51%; p < 0.05), but there was no change in the medial pre‐optic area (mPOA; 14 vs 19%). CRFR type 2 expression in the lower part of the ARC and the median eminence was not altered by insulin treatment. Thus, disruption of the LH surge after IIH in the follicular phase is not associated with decreased kisspeptin cell activation or an increase in CRFR type 2 in the ARC but may involve other cell types located in the ARC nucleus which are activated in response to IIH.     

Intercohort cannibalism and parturition-associated behaviour of adult swordtail, Xiphophorus helleri Heckel (Pisces: Poeciliidae)

The post‐parturition behavioural patterns of adult Xiphophorus helleri (Heckel) were described and quantified by recording their duration and/or frequency in glass tanks. All births occurred in the dark. Female behaviour might increase the chance of newborn juvenile survival: at the top of the tank, where juveniles are more vulnerable to predation, the females gave birth in the artificial refuge, while no parturition took place in the refuge at the bottom. Birth‐giving females were no less cannibalistic than males and other females. There was a reduction in cannibalism and the frequency of secondary attacks with time, even though the number of juveniles in the tank continued to increase due to ongoing parturition. Most adults were found at the bottom of the water column during the first 500 min of parturition, after which they were distributed equally in the water column. The movement of adults, away from the bottom and consequently away from the juveniles corresponded with an increase in feeding on alternative food and a reduction in cannibalism. Thus, cannibalism in swordtail might be opportunistic predation.     

Large-scale turbidostat culture of marine microalgae

A simple and inexpensive device is described which maintains cultures of marine microalgae at predetermined cell densities by the automatic control of harvesting and replenishment of the culture volume with fresh medium. Cultures were maintained at a range of cell densities in 80-litre internally illuminated vessels and the daily cell production yield was evaluated.Maximum yields of 0·8–1·0×10¹² cells day^?1 of Isochrysis aff. galbana Green and 0·8–1·2×10¹¹ cells day^?1 of Tetraselmis suecica (Kylin) Butch. were obtained at culture densities of 15–17×10³ cells μlitre^?1 and 19–21×10² cells μlitre^?1, respectively. Turbidostat cultures of Skeletonema costatum (Grev.) Cleve could not consistently be maintained above 5–7×10³ cells μlitre^?1.     

A Large‐scale,Rapid Public Health Response to Rabies in an Organ Recipient and the Previously Undiagnosed Organ Donor

This article describes and contrasts the public health response to two human rabies cases: one organ recipient diagnosed within days of symptom onset and the transplant donor who was diagnosed 18 months post‐symptom onset. In response to an organ‐transplant‐related rabies case diagnosed in 2013, organ donor and recipient investigations were conducted by multiple public health agencies. Persons with potential exposure to infectious patient materials were assessed for rabies virus exposure. An exposure investigation was conducted to determine the source of the organ donor's infection. Over 100 persons from more than 20 agencies spent over 2700 h conducting contact investigations in healthcare, military and community settings. The 564 persons assessed include 417 healthcare workers [5.8% recommended for post‐exposure prophylaxis (PEP)], 96 community contacts (15.6% recommended for PEP), 30 autopsy personnel (50% recommended for PEP), and 21 other persons (4.8% recommended for PEP). Donor contacts represented 188 assessed with 20.2% recommended for PEP, compared with 5.6% of 306 recipient contacts recommended for PEP. Human rabies cases result in substantial use of public health and medical resources, especially when diagnosis is delayed. Although rare, clinicians should consider rabies in cases of encephalitis of unexplained aetiology, particularly for cases that may result in organ donation.     

Duration and method of fixation affects the sensitivity of a digoxygenin-labelled DNA probe in detecting Kudoa thyrsites in Atlantic salmon skeletal muscle

The effect of fixative and duration of fixation on the sensitivity of a non-radioactive in situ hybridisation (ISH) protocol to detect Kudoa thyrsites small subunit ribosomal deoxyribonucleic acid (DNA) was investigated. Strong ISH reactions were detected in 5-μm sections of paraffin-embedded Atlantic salmon muscle after fixation for 1 day in Davidson's solution (DS). Reactions were weak following 3 or 5 days fixation and absent after 17 or 28 days fixation. Strong ISH reactions were observed after 1, 3 or 5 days fixation in neutral buffered formalin (NBF). The reactions were weak after 17 days and weak to nonexistent after 28 days of fixation. Reactions were consistently strong after fixation in 95% ethanol for up to 28 days. Some mature spores reacted weakly or not at all by ISH. Parasite DNA was weakly amplified by polymerase chain reaction (PCR) from paraffin-embedded muscle after 1 day of fixation in DS but not after fixation for 3, 5, 17 or 28 days. Amplified DNA was detected after fixation in NBF for 1, 3 and 5 days, but not after 17 or 28 days. In contrast, PCR consistently amplified DNA from paraffin-embedded, ethanol-fixed muscle. Caution should be used in the choice of fixative and duration of fixation when preserving Atlantic salmon tissues for molecular diagnosis of K. thyrsites.     

The effect of dietary protein source on growth and carcass composition in juvenile Australian freshwater crayfish

A feed trial was conducted for 12 weeks on juvenile Australian freshwater crayfish (Cherax destructor) (mean weight (SE) 0.82 (0.02)g) maintained on five isoenergetic diets with a protein content of 30%. Diets differed in the primary source of protein used, with meat, snail, soybean, yabby, and zooplankton meals comprising the major protein ingredient, varying from 56–60% of total protein. Mean percentage weight gain per day ranged from 7.57% (yabby meal diet), to 9.42% (snail meal diet). No significant difference occurred in mean weight, percentage weight gain, specific growth rate (%), or survival among diets. A maximum size of 16.44g was attained on the yabby meal diet. Largest mean weight was 8.27g on the snail-based diet. Food conversion ratios were all good, with a minimum value of 0.95 recorded for the snail-based diet. Initial food consumption per day was approximately 5% of mean animal weight, decreasing to around 2.4%, and is collectively described by a power curve. Protein retention ranged from 29.57% in the zooplankton meal diet to 41.15% in the snail-based diet. Carcass composition was influenced by feed type, with the most marked difference occurring in carapace colour. Animals fed the zooplankton-based pellets developed the strongest pigmentation. Recommendations are made for including certain protein-based ingredients in manufactured yabby diets.