Lesion Scores Research Articles

Exploring porcine kidney explants as a model for the study of nephrotoxins and the therapeutic potential of phytic acid

The use of in vivo models to assess nephrotoxicity has faced ethical limitations. A viable alternative is the ex vivo model that combines the 3 R principles with the preservation of tissue histology. Here, we established a gentamicin nephrotoxicity model using pigs` kidney explants and investigated the effect of phytic acid (IP6) against gentamicin- induced nephrotoxicity. A total of 360 kidney explants were divided into control, gentamicin (10 mM), IP6 (5 mM), and gentamicin+IP6 groups. The activity of gammaglutamyltransferase (GGT), creatinine levels, histological assessment, oxidative stress, and inflammatory cytokine expression were analyzed. Exposure to gentamicin induced an increase in GGT activity, creatinine levels, lesion score, lipoperoxidation and IL-8 expression. Explants exposed to IP6 remained like the control. The addition of IP6 to gentamicin prevented tissue damage, increasing the antioxidant status and gene expression of IL-10. This model proved to be an adequate experimental approach for identifying nephrotoxins and potential products to modulate the toxicity.

Assessing the effects of piglet behavior, diet supplementation, and lesion score breeding values of sires on behavior pattern in weaned pigs

The aim of the present study was to study effects of feeding in terms of a herbal diet supplement (HD; basal diet plus a supplement with chicory herbs), of early piglet backtest scores and of relative breeding values for skin lesions (RBV-LS) on behavior pattern of weaned pigs during the post-weaning period. In this regard, we implemented a balanced research design, allowing a semi-randomized allocation of pigs in groups with a very similar number of observations for all sub-cells of diet x backtest score x RBV-LS class combinations. With regard to backtest scores, piglets were classified as high-resisting (HR), low-resisting (LR) or intermediate-resisting (IR). RBV-LS were estimated for the sires based on skin lesions from 993 offspring. The sires were categorized into 2 groups with RBV-LS > 100 (favorable genetic value indicating only a few or no lesions) or RBV-LS ≤ 100 (indicating a large number or severe lesions). Video images were analyzed one day after weaning and 5 weeks later, and considered 300 min per monitoring date for the 8 different possible behavior traits resting time (REST), body contact (BCON), initiating fights (IFIGHT), fighting (FIGHT), refusing of fights (RFIGHT), ear or tail biting (BITE), explorative behavior (EXPLORE) and remaining activities (RACT). Finally, 104 pigs with complete observations over the total video monitoring time at both recording dates, and with complete observations for the explanatory variables sire RBV-LS, backtest score and feeding group, were considered for ongoing association studies. The 104 pigs were crosses from matings of Piétrain boars with German Landrace or German Edelschwein sows. The 104 pigs were offspring of 9 different boars (sire lines). Effects of feeding, backtest score and RBV-LS class on video behaviors were inferred via mixed model analyses, implying 8 different runs for the 8 video behavior traits. Least squares means (lsmeans) significantly (P < 0.05) differed between HD and the control feeding group (CON) for FIGHT and BITE, with longer durations of aggressive behavior for CON pigs. Vice versa, lsmeans for REST indicating calm behavior were larger for the HD group. Pigs classified as HR piglets were more aggressive than the LR and IR contemporaries, with significantly higher lsmeans for FIGHT, BITE and EXPLORE. The pigs allocated to the sire RBV-LS > 100 group had significantly (P < 0.05) longer durations for REST and RFIGHT. Vice versa, pigs allocated to the sire RBV-LS ≤ 100 groups spent more with aggressive behavior in terms of BITE and FIGHT. Consequently, boar RBV-LS were favorably correlated with REST, IFIGHT and RACT. In conclusion, the early piglet backtest scores as well as breeding values from a victim perspective (lesion scores) can be used as indicator traits for selection against aggressiveness. Nevertheless, results from the present study are first indications and breeding value correlations are approximations in this regard, which should be validated in ongoing studies based on larger datasets via real genetic correlation estimates between actor and victim traits, and considering social interactions and interactions between all fixed effects simultaneously.

Influences of flavonoid (quercetin) inclusion to corn-soybean-gluten meal-based diet on broiler performance.

Quercetin (a predominant flavonoid) is considered to have antimicrobial and antioxidant properties. This trial was conducted to evaluate the impact of graded doses of quercetin (QS) on growth efficiency, nutrient retention, faecal score, footpad lesion score, tibia ash and meat quality. In a 32-day feeding test, a total of 576 1-day-old Ross 308 broilers (male) were allocated arbitrarily with an average body weight of 41 ± 0.5 g. The trial had four dietary treatments with eight repetitions of 18 birds per pen and a basal diet incorporating 0%, 0.02%, 0.04% and 0.06% of QS. As the QS dosage increased, body weight gain tended to increase linearly on Days 9-21 (p = 0.069) and overall period (p = 0.079). Similarly, feed intake increased (p = 0.009) linearly with the increasing doses of QS on Days 9-21. Likewise, there was a linear improvement in dry matter (p = 0.002) and energy (p = 0.016) digestibility after QS administration. Moreover, the inclusion of QS supplement (0%-0.06%) linearly increased (p = 0.012) tibia ash in broilers. However, the faecal score and footpad lesion score showed no significant outcome (p > 0.05). By giving broilers a graded amount of QS, the relative organ weights of breast muscle (p = 0.009) and spleen (p = 0.006) improved linearly, meat colour lightness increased (p = 0.015), redness tended to improve (p = 0.065) linearly and drip loss decreased (p = 0.015) linearly. The inclusion of QS in the graded-level diet led to improvements in growth efficiency, nutrient absorption, meat quality and tibia ash, which recommended it as a beneficial feed additive for the broiler.

Are T2WI PI-RADS sub-scores of transition zone prostate lesions biased by DWI information? A multi-reader, single-center study

PurposeAccording to PI-RADS v2.1, T2-weighted imaging (T2WI) is the dominant sequence for transition zone (TZ) lesions. This study aimed to assess, whether diffusion-weighted imaging (DWI) information influences the assignment of T2WI scores. MethodOut of 283 prostate MRI examinations with correlated biopsy results, fourty-four patients were selected retrospectively: first, 22 patients with a TZ lesion with T2WI and DWI scores ≥ 4, to represent lesions with unequivocal suspicion on T2WI and DWI. Second, 22 additional patients with TZ lesions of similar T2WI appearance but with corresponding DWI score ≤ 3 were added as control. Four residents and one board-certified radiologist each performed two assessments of the included patients: First, only T2WI was available (T2-only read); second, both T2WI and DWI sequences were available (biparametric read). Lesion scores were assessed using Wilcoxon signed-rank test, inter-reader agreement using weighted kappa and Kendall's W statistics, and sensitivity/specificity using McNemar test. ResultsThe T2WI scores were significantly different between the T2-only and biparametric read for 3 out of 4 residents (p ≤ 0.049) but not for the radiologist. The overall PI-RADS scores derived from the two reading sessions differed considerably for 35/220 cases (all readers pooled). Inter-reader agreement was fair for the T2WI and overall PI-RADS scores (mean kappa 0.27–0.30) and moderate for the DWI scores (mean kappa 0.43). ConclusionsFor inexperienced readers, assessment of T2WI is variable and potentially biased by availability of DWI information, which can lead to changes of overall PI-RADS score and consequently clinical management. Assessment of T2WI should be performed before reviewing DWI to ensure non-biased interpretation of TZ lesions in the dominant sequence.

Impact of Residual Lesion Severity on Neurodevelopmental Outcomes Following Congenital Heart Surgery in Infancy and Childhood.

Children with congenital heart disease are at increased risk of neurodevelopmental delay throughout their lifespan. This risk is exacerbated following congenital heart surgery (CHS) in infancy. However, there are few modifiable risk factors for postoperative neurodevelopmental delay. In this study, weassessed the Residual Lesion Score (RLS), a quality assessment metric that evaluates residual lesion severity following CHS, as a predictor of neurodevelopmental delay. This was a single-center, retrospective review of patients who underwent CHS from 01/2011 to 03/2021 and post-discharge neurodevelopmental evaluation from 12 to 42months of age using the Bayley Scales of Infant Development, 3rd Edition (BSID-III). RLS was assigned per published criteria: RLS 1, no residua; RLS 2, minor residua; and RLS 3, major residua or pre-discharge reintervention. Associations between RLS and BSID-III scores, as well as trends in neurodevelopmental outcomes over time, were evaluated. Of 517 patients with median age at neurodevelopmental testing of 20.0 (IQR 18.0-22.7) months, 304 (58.8%), 146 (28.2%), and 67 (13.0%) were RLS 1, 2, and 3, respectively. RLS 3 patients had significantly lower scaled scores in the cognitive, receptive, and expressive communication, and fine and gross motor domains, compared with RLS 1 patients. Multivariable models accounted for 21.5%-31.5% of the variation in the scaled scores, with RLS explaining 1.4-7.3% of the variation. In a subgroup analysis, RLS 3 patients demonstrated relatively fewer gains in cognitive, expressive communication, and gross motor scores over time (all p < 0.05). In conclusion,RLS 3 patients are at increased risk for neurodevelopmental delay, warranting closer follow-up and greater developmental support for cognitive, language, and motor skills soon after surgery.

Incidence of hoof and leg injuries in dairy cow under different floor

Present study explored the effect of different floor on hoof health and leg injuries of crossbred dairy cow. Animals (n=24), were randomly assigned into four groups with different floor combination in covered and open area, viz. T0 (concrete and brick paved), T1 (concrete and sand), T2 (rubber mat and sand) and T3 (rubber mat and brick paved). Hoof and limb lesion scores were recorded as per the standard protocol during study period. White line lesion score did not differ significantly, however, sole lesion score and inter-digital space lesion score differed significantly between the groups. Both, sole and interdigital space lesion scores were greater in T0 group. Total hoof lesion score was different between the groups with highest value in T0 group. Month wise hoof lesion scores differedsignificantly in 5th and 6th month between treatments. Decreasing trend was noticed in T1 and T2 from 5th month onwards. In contrast, T0 showed increase in hoof lesion score. Knee lesion score and hock lesion score were different between treatments. Higher scores were found in cows housed in T0 group, while lower scores were noted in T1 group. Knee lesion score decreased in T1 and T2 groups 4th month onwards, however no change was seen in T0 and T3 group. Similar trend was also seen for hock lesion score. In conclusion, provision of sand bed floor in open area of loose house with or without rubber mat floor in covered area was found superior for improving animal welfare in terms of lesser hoof and limb injuries.

ECoG spiking activity and signal dimension are early predictive measures of epileptogenesis in a translational mouse model of traumatic brain injury

The latency between traumatic brain injury (TBI) and the onset of epilepsy (PTE) represents an opportunity for counteracting epileptogenesis. Antiepileptogenesis trials are hampered by the lack of sensitive biomarkers that allow to enrich patient's population at-risk for PTE. We aimed to assess whether specific ECoG signals predict PTE in a clinically relevant mouse model with ∼60% epilepsy incidence. TBI was provoked in adult CD1 male mice by controlled cortical impact on the left parieto-temporal cortex, then mice were implanted with two perilesional cortical screw electrodes and two similar electrodes in the hemisphere contralateral to the lesion site. Acute seizures and spikes/sharp waves were ECoG-recorded during 1 week post-TBI. These early ECoG events were analyzed according to PTE incidence as assessed by measuring spontaneous recurrent seizures (SRS) at 5 months post-TBI. We found that incidence, number and duration of acute seizures during 3 days post-TBI were similar in PTE mice and mice not developing epilepsy (No SRS mice). Control mice with cortical electrodes (naïve, n = 5) or with electrodes and craniotomy (sham, n = 5) exhibited acute seizures but did not develop epilepsy. The daily number of spikes/sharp waves at the perilesional electrodes was increased similarly in PTE (n = 15) and No SRS (n = 8) mice vs controls (p < 0.05, n = 10) from day 2 post-injury. Differently, the daily number of spikes/sharp waves at both contralateral electrodes showed a progressive increase in PTE mice vs No SRS and control mice. In particular, spikes number was higher in PTE vs No SRS mice (p < 0.05) at 6 and 7 days post-TBI, and this measure predicted epilepsy development with high accuracy (AUC = 0.77, p = 0.03; CI 0.5830–0.9670). The cut-off value was validated in an independent cohort of TBI mice (n = 12). The daily spike number at the contralateral electrodes showed a circadian distribution in PTE mice which was not observed in No SRS mice. Analysis of non-linear dynamics at each electrode site showed changes in dimensionality during 4 days post-TBI. This measure yielded the best discrimination between PTE and No SRS mice (p < 0.01) at the cortical electrodes contralateral to injury. Data show that epileptiform activity contralateral to the lesion site has the the highest predictive value for PTE in this model reinforcing the hypothesis that the hemisphere contralateral to the lesion core may drive epileptogenic networks after TBI.

Health effects of feed dilution and roughage in Ross 308 broiler breeder cockerels

To ensure reproductivity and prevent obesity, broiler breeder's growth rate is controlled by quantitative feed restriction. However, feed restriction is associated with chronic hunger, frustration, and abnormal behaviors, thus representing a welfare problem. Feed diluted with insoluble fiber is an alternative, allowing larger amounts of feed and more gut filling, increasing satiety without increasing the caloric intake. Previous research on feed dilution has focused on pullets not cockerels. In addition, the health effects of diluted feed are less explored. The aim of this study was to investigate the effects of feed dilution and roughage on health indicators in broiler breeder cockerels, 5 to 10 wk of age (WOA), a very feed restricted period. In total, 200 Ross 308 broiler breeder cockerels were housed in 12 pens (6 pens/treatment), 16 to 17 birds per pen. The treatments were standard feed (Control) and feed diluted with (20%) insoluble oat hulls and 150 g of roughage (lucerne/alfalfa) daily per pen (D+R). The D+R birds received 20% more feed per day. Body weight, pecking injuries, footpad dermatitis (FPD), and litter quality were recorded weekly between 5 and 9 WOA. At 10 WOA all birds were euthanized and subjected to postmortem assessment. D+R birds tended to weigh more than controls (P=0.055), but diet did not affect flock uniformity (P=0.57). The weekly assessment showed no effect of treatment on foot pad lesion scores (P=0.31). However, an effect on FPD was observed postmortem (P=0.04), where Control had slightly better footpad condition compared to the D+R group. Litter had to be completely changed in 2 control pens during the trial, due to quality deterioration. There was no effect of treatment on the weight of specific organs, the length of the gut, the intestinal mucosa, nor on the incidence of wounds on the head (P=0.15). Overall, these results indicate that feed dilution and roughage have neither adverse nor beneficial health effects on Ross 308 broiler breeder cockerels.

Lactococcus lactis NZ3900/pNZ8149‐IL‐4‐IL‐2 as an adjuvant to reduce vaccine dose in chicken coccidia live mixed vaccine

AbstractAlthough live mixed coccidiosis vaccines are widely used for Eimeria control, the side effects (e.g., damaging the intestinal mucosa and decreasing their weight gain) are not negligible. This study aimed to filter out the lowest dose of coccidiosis mixed vaccine with Lactococcus lactis (L. lactis) NZ3900/pNZ8149‐IL‐4‐IL‐2 (RLIL4/2) coimmunization against Eimeria reduces the side effects for Eimeria vaccination. Chickens oral 1.0 ×, 0.9 ×, 0.8 ×, 0.6 ×, 0.4 × dose of live vaccine, and RLIL4/2 with 1.0 × 109 CFU/chicken, respectively. The blank control group, challenge‐only group and 1.0 × dose vaccine‐only group are used as references. The survival rate, weight gain, bloody stool, intestinal lesions scores, oocyst output oocysts per gramme (OPG), and anticoccidial index (ACI) were detected by challenging with virulent coccidiosis. All the results indicated that the weight gain during the immune period or bloody stool/intestinal lesions scores/OPG during the challenge period were oppositely correlated with vaccine doses. The survival rate, weight gain, and ACI were positively correlated with vaccine doses during the challenge period. The ACI of 0.6 × the dose group was 167.69, which was considered a moderate efficiency. The weight gain, ACI of 1.0 ×, 0.9 × and 0.8 × dose groups were higher or intestinal lesions were lower than 1.0 × dose vaccine‐only group (p &lt; 0.05), respectively. In conclusion, the coimmunization of RLIL4/2 with Eimeria mixed vaccine could reduce 40% of the dose and the side effects (e.g., weight gain and intestinal lesions) from live mixed coccidiosis vaccine.

Effects of a complex probiotic preparation, Fengqiang Shengtai and coccidiosis vaccine on the performance and intestinal microbiota of broilers challenged with Eimeria spp.

BackgroundCoccidiosis, a prominent intestinal protozoan disease, carries significant economic implications for the poultry industry. The aim of this study was to evaluate the effects of Fengqiang Shengtai (BLES), a probiotics product, and coccidiosis vaccine in modulating the intestinal microbiome and providing insight into mitigating the occurrence and management of avian coccidiosis.MethodsBroilers included in the study were divided into four pre-treatment groups: the Pre-Con group (commercial diet), Pre-BLES group (BLES supplement), Pre-Vac group (coccidiosis vaccination) and Pre-Vac-BLES group (combined vaccination and BLES). Body weight gain, feed consumption and feed conversion ratio were monitored from age 25 to 55 days. Cecum contents were collected at 8 and 15 days of age for comparative analysis of intestinal microbiomes. In the Pre-BLES and Pre-Vac-BLES groups, probiotics were administered at a dose of 0.01 g per chicken between ages 3 to 6 days and 10–13 days. At 3 days of age, chickens in the Pre-Vac and Pre-Vac-BLES groups were vaccinated with 1700 sporulated oocysts of the live coccidiosis vaccine per chicken. At the age of 25 days, Eimeria spp. challenge experiments were performed based on the aforementioned immunization strategy, and the oocysts per gram (OPG) in the feces, intestinal lesion score and intestinal pathological characteristics were evaluated. Specifically, 30 chickens were randomly selected from each group and orally administered 34,000 sporulated oocysts of Eimeria spp. per chicken, re-defined as Eimeria group, BLES-Eimeria group, Vac-Eimeria group and Vac-BLES-Eimeria group, respectively. Additionally, 30 chickens were randomly selected from the Pre-Con group and included as negative control without Eimeria spp. challenge. Intestinal microbiota was sequenced and analyzed when the broilers were 32 days old.ResultsA significant improvement was observed in body weight gain of the broilers in the Pre-BLES and Pre-Vac-BLES group at 45 days of age. Analysis of the intestinal microbiota revealed a positive correlation between the experimental groups receiving BLES and coccidiosis vaccines at 8 and 15 days of age with the Enterococcus genus and Lachnospiraceae NK4A136 group, respectively. In addition to the reduced lesion score and OPG values, the combination of coccidiosis vaccine and BLES also reduced the intestinal epithelial abscission induced by coccidiosis vaccines. The results of intestinal microbial function prediction demonstrated that N-glycan biosynthesis and ferroptosis were the prominent signal pathways in the Vac-BLES-Eimeria group.ConclusionsTaken together, the results of the present study suggest that supplementation of BLES with coccidiosis vaccine represents a promising strategy for improving growth performance, alleviating clinical manifestations and inducing favorable alterations to the intestinal microbiota in broiler chickens affected by coccidiosis.

Live IBD vaccine exacerbates disease and pathological effects of Asian lineage H9N2 LPAIV in chickens

ABSTRACT Avian influenza H9N2 is one of the most commonly circulating viruses in numerous Egyptian poultry farms. The Asian lineage H9N2 exhibits an immunosuppressive effect, and its pathogenicity is amplified when it co-infects with other pathogens, especially with the immunosuppressive infectious bursal disease virus (IBDV), resulting in increased mortality rates. Both vaccines and field infection can exacerbate the pathogenicity of H9N2, particularly in the bursa of Fabricius, causing more significant lymphoid depletion. To comprehend the impact of the IBD vaccine on the viral and pathogenic effect of H9N2 infection in specific pathogen-free chicks (SPF), the experiment was designed as four groups; group 1 served as the negative control, group 2 received (228E) IBD vaccine, group 3 was challenged with H9N2, and group-4 was vaccinated by the IBD vaccine then challenged with H9N2. The clinical signs, relative immune organs weights and histopathological lesion scores were recorded. The tracheal and cloacal H9N2 viral shedding were also measured. Group 4 exhibited a significant decrease (P ≤ 0.05) in the relative bursal weight and an increase in the bursal lesion score when compared with groups 1 and 3 at 4 and 8 days post-challenge (dpc). The tracheal lesion score of group-4 recorded a significant increase when compared with groups 1 and 3. The renal lesion score of group 4 achieved a significant increase when compared with 1 and 3 at 8 dpc. Also, group 4 recorded a significant increase in H9N2 shedding in comparison with groups 1 and 3. Consequently, our study concluded that routine vaccination with the IBD intermediate plus vaccine exacerbates the silent infection of H9N2 resulting in outbreaks.

Effects of doxycycline dose rate and pre-adulticide wait period on heartworm-associated pathology and adult worm mass

BackgroundThe American Heartworm Society canine guidelines recommend treatment with doxycycline prior to adulticide administration to reduce levels of Wolbachia and its associated metabolites, which are known to be a leading cause of pulmonary pathology. Studies have determined that doxycycline administered at 10 mg/kg BID for 28 days is an effective dose for eliminating Wolbachia, but what has not been determined is the clinical relevance of this elimination. The current guidelines also recommend a 30-day wait period following administration of doxycycline to allow for clearance of metabolites, such as Wolbachia surface protein, and for further reduction in heartworm biomass before administration of adulticide. Reducing the doxycycline dose and eliminating the wait period may carry practical benefits for the animal, client, and practitioner.MethodsTo investigate these treatment practices, Dirofilaria immitis adults were surgically transplanted into each of 45 dogs, which were divided into nine study groups of five dogs each. Seventy-five days after transplantation, two groups each were administered 5, 7.5, or 10 mg/kg BID doxycycline orally for 28 days and 6 µg/kg ivermectin monthly, with three untreated groups serving as controls. Study animals were necropsied and examined prior to treatment as well as 30 and 60 days post-treatment.ResultsMean worm weight was unaffected by dosage but exhibited a significant increase at 30 days and significant decrease at 60 days post-treatment, including in control groups. Histopathology lesion scores did not significantly differ among groups, with the exception of the lung composite score for one untreated group. Liver enzymes, the levels of which are a concern in doxycycline treatment, were also examined, with no abnormalities in alanine aminotransferase or alkaline phosphatase observed.ConclusionsNo consistent worsening of tissue lesions was observed with or without the AHS-recommended 30-day wait period, nor did reduced dosages of doxycycline lead to worsening of pathology or any change in efficacy in depleting worm weight. Mean worm weight did significantly increase prior to, and decrease following, the wait period. Future work that also includes adulticide treatment (i.e. melarsomine) will study treatment recommendations that may improve both animal health and owner compliance.Graphical abstract

In Vitro and In Vivo Evaluation of Composite Oral Fast Disintegrating Film: An Innovative Strategy for the Codelivery of Ranitidine HCl and Flurbiprofen.

Here, we evaluate the feasibility of co-loading plain ranitidine hydrochloride (RHCl) and microencapsulated flurbiprofen (FBP) in a Lycoat® RS780-based oral fast disintegrating film (ODF). These films were developed by the solvent casting method to minimize the adverse effects of FBP and reduce the dosage form burden on patients. Optimized FBP microparticles (M3) with an average size of 21.2 ± 9.2 µm were loaded alone (F1) and in combination with plain RHCl (F2) in the composite ODF. All films were evaluated physicomechanically and physicochemically. These films were resilient, flexible, and disintegrated within thirty seconds. SEM images showed intact FBP microparticles in both formulations and, moreover, did not observe an interaction between the drug and film components. Microencapsulated FBP was released in a controlled manner over 48 h from the proposed formulations, while RHCl was released within 5 min from F2. After in vitro evaluation, formulations were also tested for in vivo anti-inflammatory activity, cytokine (TNF-α and IL-6) levels, and gastroprotective effects in rats. The anti-inflammatory activity and gastroprotective effect of F2 were markedly higher than pure FBP and other synthesized formulations (M3 and F1). The average score of gastric lesions was in the order of pure FBP (15.5 ± 1.32) > M3 (8 ± 2) > F1 (1 ± 0.5) > F2 (0.5 ± 0) > control (0). Additionally, F2 showed a sustained anti-inflammatory effect up to 10 h in the rat paw edema model. Furthermore, F2 also markedly reduced TNF-α and IL-6 levels. Conclusively, the Lycoat® RS780-based composite film could be a promising carrier for the co-loading of microencapsulated FBP with RHCl. In the future, an optimized formulation (F2) could be capable of countering the issues related to multiple drug administration in geriatric patients and evading the gastric irritation associated with FBP.

Protective efficacy of recombinant proteins AMA1 and IMP1 in rabbits infected with Eimeria intestinalis

Eimeria intestinalis is one of the most pathogenic rabbit coccidia species causing severe intestinal damage and increased risk of secondary infection from opportunistic pathogens, which results in huge economic losses to the rabbit industry. Anticoccidial drugs are currently the main method to control coccidiosis; however, increasing resistance and drug residues have fueled research on anticoccidial vaccines. Apical membrane antigen 1 (AMA1) and immune mapped protein 1 (IMP1), as surface proteins, are associated with host invasion and might have the potential as candidate vaccine antigens. In the present study, recombinant IMP1 (rEiIMP1) and AMA1 (rEiAMA1) from E. intestinalis were expressed using Escherichia coli BL21. The immunoreactivity and immunoprotective effects of rEiIMP1 and rEiAMA1 were then analyzed. Fifty rabbits were grouped randomly (n = 10 per group): The unimmunized-unchallenged control group (sterilized phosphate-buffered saline (PBS)), the unimmunized-challenged control group (sterilized PBS), the vector protein-challenged control group (100 μg of pET-32a vector protein per rabbit), the rEiIMP1 immunized group (100 μg of rEiIMP1 per rabbit), and the rEiAMA1 immunized group (100 μg of rEiAMA1 per rabbit). After two immunizations, the rabbits were challenged with homologous oocysts (except for the unimmunized-unchallenged group). Serum specific antibody levels were assessed weekly throughout the experimental period; and the levels of different cytokines in the serum before the challenge were detected. The clinical symptoms, oocysts output, weight gain, feed conversion ratio (FCR), and lesion scores were recorded after experimental infection, and the anticoccidial indexes (ACIs) were calculated. The results showed that both rEiIMP1 and rEiAMA1 had good immunoreactivity. Rabbits immunized with rEiIMP1 and rEiAMA1 displayed 66.74 % and 63.14 % oocyst reduction, respective land 81.79 % and 78.87 % body weight gain, respectively. The rEiIMP1 and rEiAMA1 groups had lower FCRs (3.77:1 and 4.06:1, respectively) and lesion scores (P = 0.00). The rEiIMP1 and rEiAMA1 showed moderate effects, with an ACI of 152.09 and 147.17, respectively. Immunization induced high levels of anti-rEiIMP1 and -rEiAMA1 antibodies. Rabbits immunized with rEiIMP1 and rEiAMA1 displayed significantly increased interleukin (IL)− 2 (P = 0.00), interferon gamma (IFN)− γ (P = 0.00), and IL− 4 (P = 0.00) levels. Therefore, this study provided potential candidate vaccine antigens for E. intestinalis.

Lactobacillus plantarum surface-displayed Eimeria tenella profilin antigens with FliC flagellin elicit protection against coccidiosis in chickens

Coccidiosis is a parasitic disease in the intestine caused by the genus Eimeria that poses a substantial economic threat to the broiler breeding industry. The misuse of chemoprophylaxis and live oocyst vaccines has a negative impact on chicken reproductivity. Therefore, there is a pressing need to develop safe, convenient, and effective vaccines. Lactic acid bacteria can be used as a means to deliver mucosal vaccines against intestinal pathogens, which is a promising strategy. In this study, a recombinant Lactobacillus plantarum (L. plantarum) with surface-expressed antigens constructed from the fusion of Eimeria tenella (E. tenella) antigen profilin and the Salmonella enterica serovar Typhimurium flagellin protein FliC was created. After oral immunization with the recombinant L. plantarum, T-cell differentiation was analyzed by flow cytometry, and specific antibody levels were determined via indirect ELISA. Oocyst shedding, body weight, and cecum lesions were assessed as measures of protective immunity after challenge with E. tenella. The results of this study demonstrate the effectiveness of recombinant L. plantarum as an immunization agent for chickens. Specific IgA titers in the intestine and specific IgG antibody titers in the serum were significantly higher in chickens immunized with recombinant L. plantarum (P < 0.001). Additionally, the levels of IL-2 (P < 0.05) and IFN-γ (P < 0.01) in the serum were markedly increased. Recombinant L. plantarum induced T-cell differentiation, resulting in a higher proportion of CD4+ and CD8+ T cells in splenocytes (P < 0.001). Fecal oocyst shedding in the immunized group was significantly reduced (P < 0.001). Additionally, recombinant L. plantarum significantly relieved pathological damage in the cecum, as evidenced by lesion scores (P < 0.01) and histopathological cecum sections. In conclusion, the present study provides evidence to support the possibility of using L. plantarum as a promising carrier for the delivery of protective antigens to effectively protect chickens against coccidiosis.

A novel organotypic cortical slice culture model for traumatic brain injury: molecular changes induced by injury and mesenchymal stromal cell secretome treatment.

Traumatic brain injury (TBI) is a major worldwide neurological disorder with no neuroprotective treatment available. Three-dimensional (3D) in vitro models of brain contusion serving as a screening platform for drug testing are lacking. Here we developed a new in vitro model of brain contusion on organotypic cortical brain slices and tested its responsiveness to mesenchymal stromal cell (MSC) derived secretome. A focal TBI was induced on organotypic slices by an electromagnetic impactor. Compared to control condition, a temporal increase in cell death was observed after TBI by propidium iodide incorporation and lactate dehydrogenase release assays up to 48 h post-injury. TBI induced gross neuronal loss in the lesion core, with disruption of neuronal arborizations measured by microtubule-associated protein-2 (MAP-2) immunostaining and associated with MAP-2 gene down-regulation. Neuronal damage was confirmed by increased levels of neurofilament light chain (NfL), microtubule associated protein (Tau) and ubiquitin C-terminal hydrolase L1 (UCH-L1) released into the culture medium 48 h after TBI. We detected glial activation with microglia cells acquiring an amoeboid shape with less ramified morphology in the contusion core. MSC-secretome treatment, delivered 1 h post-injury, reduced cell death in the contusion core, decreased NfL release in the culture media, promoted neuronal reorganization and improved microglia survival/activation. Our 3D in vitro model of brain contusion recapitulates key features of TBI pathology. We showed protective effects of MSC-secretome, suggesting the model stands as a tractable medium/high throughput, ethically viable, and pathomimetic biological asset for testing new cell-based therapies.

Identification and Characterization of α-Actinin 1 of Histomonas meleagridis and Its Potential Vaccine Candidates against Histomonosis.

Histomonas meleagridis is a protozoan parasite that causes histomonosis in gallinaceous birds such as turkeys and chickens. Since the banning and restricted usage of effective drugs such as nitarsone, 80-100% morbidity and mortality occur in turkeys and 20-30% mortality in chickens. New ideas are needed to resolve the re-emergence of histomonosis in poultry. In this study, the α-actinin encoding gene from H. meleagridis was cloned. The 1839-bp gene encoding 612 amnio acids showed close phylogenetic relationships with Trichomonas vaginalis and Tritrichomonas foetus. It was then inserted into the prokaryotic expression vector pET28a(+) and induced with isopropyl-β-D-thiogalactopyranoside. A 73 kDa recombinant protein rHmα-actinin 1 was obtained and purified with a Ni-NTA chromatography column. rHmα-actinin 1 was recognized by mouse anti-rHmα-actinin 1 polyclonal antibody, mouse anti-rHmα-actinin 1 monoclonal antibody, and rehabilitation sera from H. meleagridis infected chickens. Native α-actinin 1 in the total proteins of H. meleagridis can also be detected with mouse anti-rHmα-actinin monoclonal antibody. Immunolocalization assays showed that Hmα-actinin 1 was mainly distributed in the cytoplasm of virulent histomonads JSYZ-D9 and in the peripheral regions (near the plasma membrane) of attenuated histomonads JSYZ-D195. Based on in vivo experiment, when chickens were subcutaneously immunized with rHmα-actinin 1 at 5 and 12 days old and then challenged with H. meleagridis at 19 days old, rHmα-actinin 1 reduced the lesion scores 12 days after infection (31 days old) and increased the body weight gain during the challenged period (19-31 days old). Furthermore, it also strengthened the cellular and humoral immune responses 7 days after the second immunization (19 days old). In conclusion, Hmα-actinin 1 could be used as a candidate antigen to develop vaccines against chicken histomonosis.

Understanding the Role of the Glial Scar through the Depletion of Glial Cells after Spinal Cord Injury.

Spinal cord injury (SCI) is a condition that affects between 8.8 and 246 people in a million and, unlike many other neurological disorders, it affects mostly young people, causing deficits in sensory, motor, and autonomic functions. Promoting the regrowth of axons is one of the most important goals for the neurological recovery of patients after SCI, but it is also one of the most challenging goals. A key event after SCI is the formation of a glial scar around the lesion core, mainly comprised of astrocytes, NG2+-glia, and microglia. Traditionally, the glial scar has been regarded as detrimental to recovery because it may act as a physical barrier to axon regrowth and release various inhibitory factors. However, more and more evidence now suggests that the glial scar is beneficial for the surrounding spared tissue after SCI. Here, we review experimental studies that used genetic and pharmacological approaches to ablate specific populations of glial cells in rodent models of SCI in order to understand their functional role. The studies showed that ablation of either astrocytes, NG2+-glia, or microglia might result in disorganization of the glial scar, increased inflammation, extended tissue degeneration, and impaired recovery after SCI. Hence, glial cells and glial scars appear as important beneficial players after SCI.

Evaluation of Bursal Index and Bursal Lesion Scores in Broiler Flocks

Background and Aims: The bursa of Fabricius (BF) is recognized as a vital organ that plays a crucial role in birds immunity. In order to determine the immunological status of the birds, this study was conducted to evaluate the bursal index, and bursal lesion scores in broilers.Materials and Methods: Therefore, a total of 200 broilers, 35-45 days of age, from 10 broiler flocks were randomly selected, weighed, then sacrificed. At necropsy, the bursa of Fabricius were obtained, examined, and weighed. Bursal index (BI) was calculated. Representative pieces of tissues from bursa of Fabricius were preserved in 10% neutral buffer formaldehyde for further histopathological evaluation.Results: The results obtained from (BI) calculated during the study period for all 22 broiler farms, revealed that more than 81% of the birds were included in bad group and less than 19% in mediocre group. Degree of microscopic lesions between different groups and even within one group varied from 0 (normal) to 5 (atrophy of BF) and included lymphoid necrosis in follicles, hyperplasia and folding of bursal inner epithelium, and formation of fibrous connective tissue within the follicles. Average bursal lesion scores for all groups under study was recorded as 2.09%.Conclusions: On the basis of all results including (BI) and bursal lesion scores it is concluded that bursal lymphoid structures suffer various degree of pathological lesions indicating suppressed immunity in broiler farms along with poor performance.

Transplantation of fibrin-thrombin encapsulated human induced neural stem cells promotes functional recovery of spinal cord injury rats through modulation of the microenvironment.

Recent studies have mostly focused on engraftment of cells at the lesioned spinal cord, with the expectation that differentiated neurons facilitate recovery. Only a few studies have attempted to use transplanted cells and/or biomaterials as major modulators of the spinal cord injury microenvironment. Here, we aimed to investigate the role of microenvironment modulation by cell graft on functional recovery after spinal cord injury. Induced neural stem cells reprogrammed from human peripheral blood mononuclear cells, and/or thrombin plus fibrinogen, were transplanted into the lesion site of an immunosuppressed rat spinal cord injury model. Basso, Beattie and Bresnahan score, electrophysiological function, and immunofluorescence/histological analyses showed that transplantation facilitates motor and electrophysiological function, reduces lesion volume, and promotes axonal neurofilament expression at the lesion core. Examination of the graft and niche components revealed that although the graft only survived for a relatively short period (up to 15 days), it still had a crucial impact on the microenvironment. Altogether, induced neural stem cells and human fibrin reduced the number of infiltrated immune cells, biased microglia towards a regenerative M2 phenotype, and changed the cytokine expression profile at the lesion site. Graft-induced changes of the microenvironment during the acute and subacute stages might have disrupted the inflammatory cascade chain reactions, which may have exerted a long-term impact on the functional recovery of spinal cord injury rats.