Flow cytometric detection of alpha-1-acid glycoprotein on feline circulating leucocytes

To assess whether alpha‐1‐acid glycoprotein (AGP) can be detected on the membrane of feline circulating leucocytes. Design The presence of AGP on circulating leucocytes was investigated in both clinically healthy cats and cats with different diseases. A group of feline coronavirus (FCoV)‐positive cats, comprising cats with feline infectious peritonitis (FIP) and cats not affected by FIP but seropositive for FCoV, were included in this study because the serum concentration of AGP increases during FCoV infection. Procedure Flow cytometry (using an anti‐feline AGP antibody), serum protein electrophoresis, routine haematology and measurement of the serum AGP concentration were performed using blood samples from 32 healthy cats (19 FCoV‐seropositive), 13 cats with FIP and 12 with other diseases (6 FCoV‐seropositive). The proportion of cats with AGP‐positive leucocytes in the different groups (e.g. controls vs sick; FIP vs other diseases, etc.) or in cats with different intensities of inflammatory response was compared using a Chi‐square test. Results AGP‐positive leucocytes were found in 23% of cats. Compared with controls, the proportion of patients with positive granulocytes and monocytes was higher among sick cats (especially cats with diseases other than FIP) and cats with high serum AGP concentration, but not in cats with leucocytosis or that were FCoV‐seropositive. Conclusion AGP‐positive leucocytes can be found in feline blood, especially during inflammation. Conversely, no association between AGP‐positive leucocytes and FIP was found. Further studies are needed to elucidate the mechanism responsible for this finding and its diagnostic role in cats with inflammation.     

Contrast-enhanced ultrasonography in the differentiation of retinal detachment and vitreous membrane in dogs and cats

Objectives : To evaluate contrast‐enhanced ultrasonography (CEU) and colour Doppler imaging (CDI) for detection of persistent vascularisation in retinal detachment. Methods : In 22 eyes, retinal detachment (n=13) and vitreous membranes (n=9) were confirmed by ophthalmological examination, during cataract surgery, by histopathology or after vitreoretinal surgery. Tentative diagnosis of retinal detachment or vitreous membrane was made using grey‐scale B‐mode ultrasonography. Assessment of retinal detachment was based on the presence or absence of vascularisation in the membranous structure using CDI and CEU. Results : Sensitivity, specificity, positive‐predictive value and negative‐predictive value of grey‐scale ultrasonography in differentiating retinal detachment from vitreous membrane were 92·3%, 66·6%, 80% and 85·7%. In 91% of eyes, colour Doppler assessment was unsuccessful due to the movement of the eye. Persistent vascularisation was demonstrated in all cases of retinal detachments with CEU. CEU was 100% accurate for detection and differentiation between retinal detachment and vitreous membrane. Clinical Significance : CEU is a useful clinical tool for the diagnosis of retinal detachment and vitreous membrane in dogs and cats.     

An immunoturbidimetric assay for rapid quantitative measurement of feline alpha-1-acid glycoprotein in serum and peritoneal fluid

BACKGROUND: Alpha-1-acid glycoprotein (AGP) is an acute phase protein that increases in concentration in infectious and inflammatory conditions. The serum and peritoneal fluid concentrations of AGP may be useful in the diagnosis of feline infectious peritonitis (FIP), a lethal disease of cats. Currently AGP can be measured by radioimmunodiffusion (RID) assays, which are time consuming and difficult. OBJECTIVES: The objectives of this study were to develop a rapid immunoturbidimetric assay for measurement of AGP in feline serum and peritoneal fluid and to compare the results with those obtained by RID. METHODS: AGP was purified by perchloric acid precipitation and ion-exchange chromatography from a pool of peritoneal fluid obtained from cats with FIP, as determined by a panel of laboratory tests, including serum AGP concentration, albumin: globulin ratio, and total protein concentration, anti-coronavirus antibody titers, and effusion analysis. The purified AGP in a complete Freund's adjuvant and Tween 20 mixture was injected into a sheep and blood was collected at monthly intervals. Anti-AGP antiserum, as confirmed by ELISA and Western blot techniques, and a pool of peritoneal fluid from cats with FIP were used to prepare standards. Clinical samples of feline peritoneal fluid (n=55) and serum (n=59) were assayed for AGP and results from the immunoturbidimetric and RID methods were compared. RESULTS: Significant correlation (P < .001) was obtained between methods for both peritoneal fluid (R2=.9259) and serum (R2=.9448) samples. Coefficients of variation for the immunoturbidimetric method were <5%. CONCLUSIONS: This rapid immunoturbidimetric assay for measurement of feline AGP in serum and peritoneal fluid may be of value in the diagnosis of FIP and possibly other inflammatory diseases in cats.     

Histological examination of the intestine from dogs and cats with intussusception

Objectives : To review the histological findings in the intestine from dogs and cats with intussusception. Methods : Medical records and histopathology reports of dogs and cats with intussusception were reviewed retrospectively. Results : Fourty‐nine animals (31 dogs and 18 cats) were identified for inclusion. Tissues examined com‐prised the intussusception alone in 29 animals (16 dogs and 13 cats), and the intussusception with additional intestinal biopsies in 20 animals (15 dogs and 5 cats). Twenty‐eight of 49 (57·1%) animals, comprising 19 of 31 (61·3%) dogs and 9 of 18 cats (50%) had abnormalities detected on histological examination of tissue. Eleven of 29 (46·9%) cases where only the intussusception was submitted achieved a histological diagnosis, compared to 17 of 20 (85%) where additional biopsies were submitted (P=0·003). Cats (median age 36 months, range 2 to 174) were significantly older than dogs (median age 7·5 months, range 1 to 125 months, P=0·010) and were significantly more likely to have underlying neoplasia (5 of 9; 55·6%) compared to dogs who were more likely to have inflammatory causes (17 of 19; 89·5%, P=0·020). There was no association between histological diagnosis and location of the intussusception (P=1·000). Clinical Significance : Histological abnormalities were detected in more than half of the animals. Diagnosis of intestinal disease in animals with intussusception may be improved by submission of additional biopsy samples. Cats with intussusception are more likely to be older and have underlying neoplasia than dogs which are more likely to have inflammatory disease.     

Influence of signalment on developing cranial cruciate rupture in dogs in the UK

Objectives : To investigate risk factors associated with cranial cruciate ligament rupture in dogs. Methods : Retrospective case‐control study: medical records of a first‐opinion veterinary practice were searched for dogs diagnosed with cranial cruciate ligament rupture (1995 to 2007). For each case, six unaffected dogs were randomly selected from all dogs presenting that day for comparison. Multi‐variable binary logistic regression was performed to assess the association of variables on likelihood of cruciate rupture. Results : Frequency of cranial cruciate ligament rupture was 1·19% [95% confidence interval (CI) 1·02 to 1.36%]. West Highland white terriers (n=17), Yorkshire terriers (n=14) and Rottweilers (n=11) were at significantly increased risk of cranial cruciate ligament rupture (P≤0·002). Rottweilers were at five times greater risk compared with other pure breeds (OR 5·12, 95% CI 2·281 to 11·494, P<0·001), obesity quadrupled the risk of cranial cruciate ligament rupture (OR 3·756, 95% CI 1·659 to 8·502, P=0·001) and females were twice as likely to suffer cranial cruciate ligament failure compared to males (OR 2·054, 95% CI 1·467 to 2·877, P<0·001). Dogs less than two years old were statistically less likely to sustain cranial cruciate ligament rupture than dogs older than eight years (OR 0·246, 95% CI 0·127 to 0·477, P<0·001). There was no significant difference in median weights (in kilograms) of neutered dogs, compared to their entire counterparts in either the case group (P=0·994) or in the control group (P=0·630). There was also no significant difference in body condition (‐underweight/normal weight/overweight/obese) of neutered versus entire dogs among the cases (P=0·243), or the controls (P=0·211). Clinical Significance : Cranial cruciate ligament rupture is more likely in Rottweilers and in female dogs, older dogs and obese dogs. Following multi‐variable analysis, it was established that neutering was not associated with increased risk of cranial cruciate ligament rupture.     

Changes in some acute phase protein and immunoglobulin concentrations in cats affected by feline infectious peritonitis or exposed to feline coronavirus infection

The possible role of some acute phase proteins (APPs) and immunoglobulins in both the pathogenesis and diagnosis of feline infectious peritonitis (FIP) has been investigated. Serum protein electrophoresis and the concentration of haptoglobin (Hp), serum amyloid A (SAA), alpha(1)-acid glycoprotein (AGP), IgG and IgM were evaluated in cats exposed to feline coronavirus (FCoV) and in cats with FIP. The highest concentration of APPs was detected in affected cats, confirming the role of these proteins in supporting a clinical diagnosis of FIP. Repeated samplings from both FIP affected and FCoV-exposed cats showed that when FIP appeared in the group, all the cats had increased APP levels. This increase persisted only in cats that developed FIP (in spite of a decrease in alpha(2)-globulins) but it was only transient in FCoV-exposed cats, in which a long lasting increase in alpha(2)-globulins was observed. These results suggest that changes in the electrophoretic motility of APPs or APPs other than Hp, SAA and AGP might be involved in the pathogenesis of FIP or in protecting cats from the disease.     

Total sialic acid: An acute phase reactant in cats with a possible role in feline coronavirus infection

The aims of this study were to validate a colorimetric method to measure total sialic acid (TSA) in feline serum and to investigate the serum concentration of TSA in clinically healthy cats seronegative (n = 9) and seropositive (n = 48) for feline coronavirus (FCoV), and in cats affected by feline infectious peritonitis (FIP, n = 28), tumors (n = 20), or inflammation (n = 16). The correlation between TSA and α₁-acid glycoprotein (AGP) was also investigated. The method employed in this study is precise and accurate at TSA levels (in mg/L) commonly encountered in feline serum. No significant differences between seropositive (385.6 ± 192.2 mg/L) and seronegative (433.5 ± 179.0 mg/L) cats were detectable, suggesting that the simple infection by FCoVs does not influence TSA levels. Compared with seropositive controls, the concentration of TSA was higher in cats with FIP (556.7 ± 268.3 mg/L, P = 0.003), tumors (522.5 ± 294.4 mg/L, P = 0.028), and inflammation (546.8 ± 208.3 mg/L, P = 0.018). The discriminating power of TSA for FIP is moderate (area under the ROC curve = 0.65) and the likelihood ratio is higher than 3.0 only at high TSA levels. Consequently, TSA could support a diagnosis of FIP only at extremely high serum concentration (> 800 mg/L) or when the pre-test probability of FIP is high. No correlations were found between the TSA and AGP concentrations in cats with FIP, suggesting that sialylated proteins other than AGP are present. Both the antibody titre and the degree of AGP sialylation were negatively correlated with TSA levels, suggesting that increased TSA may contribute to reduce the burden of FCoVs.     

Seronegative conversion in four Neospora caninum-infected cows, with a low rate of transplacental transmission

Four Neospora-seropositive pregnant cows (prebreeding indirect fluorescent antibody (IFA) titers between 1:400 and 1:1600) were confined and observed until parturition. All cows gave birth to normal calves. Selected tissues were tested for NC by histopathology, immunohistochemical (IHC) and polymerase chain reaction (PCR). Parasite isolation was attempted in vero cell cultures. At parturition, all cows were seronegative at 1:200 and two of four cows had a titer of 1:100 when further tested. Three of four calves were not infected, as determined by negative results of precolostral serology (1:25 cut-off), histopathology, IHC and PCR. One calf was congenitally infected, as shown by the presence of a thick-walled cyst labelled by IHC in its brain, positive PCR of brain and a precolostral IFA titer of 1:100. It was concluded that NC antibody titers may drop or convert to seronegative status in chronically infected cows by the time of parturition and this finding in four of four cows indicates that this could be a common occurrence. Similarly, the finding of an infected calf with a low antibody titer indicates that precolostral serology may not be a fool-proof means of identifying calves with congenital Neospora caninum infections. These findings call into question conclusions of other studies that have estimated rates of congenital transmission of this parasite based on serological tests at calving. This study is the first confirmed report of congenital NC infection in a calf in Thailand.     

Comparisons between cats with normal and increased fPLI concentrations in cats diagnosed with inflammatory bowel disease

Objectives : The aim of this study was to compare age, serum albumin and cobalamin concentrations, serum alanine amino transferase and alkaline phosphatase activities, feline inflammatory bowel disease clinical disease activity index, pancreatic ultrasound findings, intestinal histopathology scores, outcome, treatment and clinical response between cats diagnosed with inflammatory bowel disease with normal or increased serum feline pancreatic lipase immunoreactivity concentrations. Methods : Medical records for 23 cats diagnosed with inflammatory bowel disease and with serum feline pancreatic lipase immunoreactivity concentrations available were reviewed. Three groups were compared; cats with serum feline pancreatic lipase immunoreactivity concentrations 2·0 to 6·8 µg/l (group A), 6·9 to 11·9 µg/l (group B) and ≥12·0 µg/l (group C). Results : Sixteen of the 23 cats had increased serum feline pancreatic lipase immunoreactivity concentrations; 9 cats in group B and 7 cats in group C. The remaining seven cats were in group A. Cats with serum feline pancreatic lipase immunoreactivity concentrations ≥12·0 µg/l had significantly lower median serum albumin and cobalamin concentrations. No significant differences were identified between the three groups for age, serum alanine amino transferase and alkaline phosphatase activities, feline inflammatory bowel disease clinical disease activity index, pancreatic ultrasound findings, intestinal histopathology scores, clinical outcome, treatment or clinical response. Clinical Significance : Hypoalbuminaemia and hypocobalaminaemia were more frequently observed in cats with serum feline pancreatic lipase immunoreactivity concentrations ≥12·0 µg/l.     

Decreased sialylation of the acute phase protein alpha1-acid glycoprotein in feline infectious peritonitis (FIP)

Feline infectious peritonitis (FIP) is an immune-mediated disease of domestic and exotic felides infected with feline coronavirus. FIP is characterized by the overexpression of an acute phase protein, the alpha1-acid glycoprotein (AGP). In humans, AGP is a heavily glycosylated protein that undergoes several modifications of its glycan moiety during acute and chronic inflammatory pathologies. We studied the changes in AGP glycosylation in the course of FIP. Specifically, we focussed our attention on the degree of sialylation, fucosylation and branching. This study presents a purification method for feline AGP (fAGP) from serum, using an ion exchange chromatography strategy. The glycosylation pattern was analyzed in detail by means of interaction of purified fAGP with specific lectins. In particular, Sambucus nigra agglutinin I and Maackia amurensis agglutinin lectins were used to detect sialic acid residues, Aleuria aurantia lectin was used to detect L-fucose residues and Concanavalin A was used to evaluate the branching degree. By this method we showed that fAGP did not present any L-fucose residues on its surface, and that its branching degree was very low, both in normal and in pathological conditions. In contrast, during FIP disease, fAGP underwent several modifications in the sialic acid content, including decreased expression of both alpha(2-6)-linked and alpha(2-3)-linked sialic acid (76 and 44%, respectively when compared to non-pathological feline AGP).     

Serum α-1-acid glycoprotein concentrations in 26 dogs with pyometra

Background: The acute phase protein, α‐1‐acid glycoprotein (AGP), has been proposed to have a role in immunomodulation and to be a nonspecific antimicrobial agent. We suggest that AGP may be increased in dogs with pyometra and possibly to a greater extent in dogs also manifesting signs of systemic inflammatory response syndrome (SIRS). Objectives: Our objectives were to evaluate serum AGP concentrations in dogs diagnosed with pyometra compared with clinically healthy female dogs and to determine if AGP concentrations were correlated with severity of disease. Methods: Twenty‐six dogs with pyometra and 18 clinically healthy intact female dogs were included in this prospective study. A diagnosis of pyometra was verified by histopathologic examination after ovariohysterectomy in the pyometra group. A commercially available single radial immunodiffusion test was used for AGP analysis. Clinical findings, laboratory variables, and hospitalization times were compared. Results: Mean AGP concentration in dogs with pyometra (1943 ± 913 mg/L, mean ± SD), was significantly higher (P<.001) than in healthy dogs (495 ± 204 mg/L). Mean AGP concentration in dogs in the pyometra group with (n=18) or without (n=8) SIRS did not differ. Animals with a prolonged hospital stay had higher AGP concentrations. Conclusions: Pyometra was associated with increased serum concentrations of the acute phase protein AGP. AGP concentrations were associated with severity of disease as measured by duration of hospitalization. As AGP binds basic drugs, further studies of its pharmacokinetic and pharmacodynamic propreties in cases of pyometra may be of clinical interest.     

Comparison of measurements of 18 analytes in canine and feline blood samples using the in-practice Falcor 350 and the reference KoneLab 30i analysers

Objectives : Falcor 350 (A. Menarini Diagnostics) is a wet‐reagent biochemistry analyser that is available for in‐house use. The aim of this study was to compare the results produced by this analyser with those obtained by a wet‐reagent analyser (KoneLab 30i; Thermo Clinical Labsystems) that served as the reference instrument. Methods : Blood samples from 120 clinical cases (60 dogs and 60 cats) were analysed for 18 analytes (urea, creatinine, total proteins, albumin, creatine kinase, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, total bilirubin, amylase, lipase, glucose, triacylglycerol, cholesterol, total calcium, phosphate, sodium and potassium) using both the reference and Falcor instruments. Results : Good to excellent correlations (r_s value) (canine/feline) were identified for urea (0·87/0·86), creatinine (0·96/0·99), total proteins (0·91/0·95), albumin (0·96/0·93), creatine kinase (0·98/0·99), aspartate aminotransferase (0·95/0·98), alanine aminotransferase (0·99/0·99), alkaline phosphatase (0·99/0·98), total bilirubin in dogs (0·88), amylase (0·99/0·87), lipase in dogs (0·88), glucose (0·97/0·98), triacylglycerol (0·93/0·97), cholesterol (0·99/0·99), total calcium (0·88/0·89), phosphate (0·94/0·97) and potassium (0·96/0·97). The correlations for sodium (0·41/0·61), total bilirubin in cats (0·78) and lipase in cats (0·25) were considered unacceptable. Clinical Significance : For 13 of the 18 analytes (creatinine, total proteins, albumin, creatine kinase, aspartate aminotransferase, alanine aminotransferase, amylase, glucose, cholesterol, triacylglycerol, phosphate, potassium and urea) in both canine and feline samples, the two instruments produce values that are closely related to each other (excellent correlation) and are sufficiently similar to allow them to be used interchangeably without the need for additional correction factor computations (good agreement). Because of differences in the methodologies, the Falcor results for alkaline phosphatase, total calcium, sodium, lipase and total bilirubin cannot be used interchangeably with those generated by the KoneLab and should be interpreted using reference intervals established from the Falcor analyser.     

Serum alpha1-acid glycoprotein (AGP) concentration in non-symptomatic cats with feline coronavirus (FCoV) infection

Previous studies have demonstrated that the concentration of alpha1-acid glycoprotein (AGP) transiently increases in asymptomatic cats infected with feline coronavirus (FCoV). In order to establish whether these fluctuations depend on the FCoV status, the serum concentration of AGP and anti-FCoV antibody titres and/or faecal shedding of FCoVs in clinically healthy cats from catteries with different levels of prevalence of FCoV infection were monitored over time. Serum AGP concentrations fluctuated over time in clinically healthy cats from the cattery with the highest prevalence of feline infectious peritonitis (FIP) and significantly increased just before an outbreak of FIP. Further studies are required to clarify whether the observed increase of AGP concentration is a consequence of the increased viral burden or a protective response against mutated viral strains. Nevertheless, the results of the present study suggest that AGP might be useful in monitoring FCoV-host interactions in FCoV-endemic catteries.     

Conventional and Doppler ultrasound for the differentiation of benign and malignant canine mammary tumours

Objectives : The aim of this study was to evaluate the sensitivity of conventional and Doppler ultrasound for differentiation of benign and malignant mammary tumours in female dogs. Methods : Mammary tumours were evaluated from 60 animals and divided into two distinct groups, group 1 (benign tumours) and group 2 (malignant tumours). The tumours were assessed by conventional ultrasound, Doppler ultrasound mode, histopathology and immunohistochemical detection of vascular endothelial growth factor. Results : Conventional ultrasound examination was found to be ineffective in separating tumours into the two experimental groups. Similarly, using colour‐flow Doppler ultrasound, no correlation was found between the presence of vascularisation and its characteristics between the two groups. Triplex Doppler ultrasound yielded average maximum velocities of 28·71 cm/s for malignant and 19·91 cm/s for benign tumours, which were significantly different (P=0·01). For vascular endothelial growth factor, an average score of 2·22 was found for group 2 and 1·66 for group 1 (P=0·03). Positive correlations were found between vascular endothelial growth factor and presence of vascularisation (P=0·04 and r=0·3658) and between vascular endothelial growth factor and maximum velocity (P=0·03 and r=0·3913). Clinical Significance : Doppler evaluation may be used to predict malignancy of mammary tumours in bitches.     

Comparison of two shampoos for the treatment of canine Malassezia dermatitis: a randomised controlled trial

Objectives : Two antimicrobial shampoos for treatment of canine Malassezia dermatitis (CMD) were compared in a prospective, randomised, single‐blinded, field clinical trial. Methods : Sixty‐seven dogs with pedal or generalised dermatitis associated with Malassezia overgrowth (MO) were treated with 3% chlorhexidine shampoo (3%CHX) or 2% miconazole‐2% chlorhexidine shampoo (2%MIC/CHX) and evaluated for up to 6 weeks until cytological recovery. Pruritus, erythema, papules, greasy seborrhoea, scaling, malodour, excoriations, secondary hairloss, lichenification, hyperpigmentation and lesion extent were each scored on a 0‐3 severity scale and combined making an aggregate score. Results : Among 54 dogs with good treatment compliance, reduction of yeast counts by at least 88% was recorded in 21 of 22 dogs with 3%CHX and 30 of 32 dogs with 2%MIC/CHX. No significant difference was detected between products for yeast count reduction (P=0·592). Time to cytological recovery was not significantly different between groups (P=0·960). Lesion score was significantly reduced in both groups after treatment (72·5 ±25·7% with 3%CHX versus 78·7 ±22·3% with 2%MIC/CHX, P=0·309). Four dogs treated with chlorhexidine shampoo showed minor adverse effects. Clinical Significance : In this study, 3%CHX was clinically as effective as 2%MIC/CHX for treatment of CMD.     

Energy and nitrogen metabolism of broilers selected over ten generations for increased growth rate,food consumption and conversion of food to gain

1. Energy and nitrogen (N) metabolism were studied in 6‐week‐old male birds taken from 4 lines of chickens selected for 10 generations for increased weight gain (line W), increased food consumption (line F), increased conversion of food to gain (line E) or at random (controls, line C).

2. Calorimetric measurements were made 8 times on each line while fed ad libitum in large open‐circuit respiration chambers for 3 d, and 11 to 13 times without food in smaller closed‐circuit respiration chambers for 24 h.

3. The F line ate 60% more food, produced 90% more excreta and 34% more heat and retained 80% more energy and 35% more N in their bodies than lines E and C. Line W was intermediate.

4. When differences in body weight were taken into account, the E and W lines had lower heat production than the C line, while the F line ate 40% more food, produced 30% more heat and retained 70% more energy and 30% more N than the E line.

5. In lines W, F, E and C respectively, the mean metabolisability of dietary energy (%) was 69·4, 62·9, 70·1 and 67·8 ; the fasting heat production (mean ± SE) was 481 ±9, 569 ± 10, 485 ± 9, and 508 ± 9 kJ/kgW d; the net availability of metabolisable energy (NAME) was 0·68 ± 0·05, 0·76 ± 0·04, 0·85 ± 0·06 and 0·73 ± 0·04; the estimated daily maintenance energy requirements were 671 + 15, 866 ± 14, 701 ± 13, and 742 ± 11 kJ ME/ kgW; and the proportion of N retained per unit increase in N intake was 0·38 ± 0·08, 0·50 ± 0·06, 0·56 ± 0·10 and 0·53 ± 0·06. 6. The contribution of line differences in the above traits to large line differences in efficiency of food utilisation is discussed.

     

Tissue distribution of a feline AGP related protein (fAGPrP) in cats with feline infectious peritonitis (FIP)

Feline alpha(1)-acid glycoprotein (fAGP) increases during feline infectious peritonitis (FIP). We have recently identified a 29 kDa protein that we named feline AGP-related protein (fAGPrP) due to its cross-reactivity with an anti-human AGP monoclonal antibody. In this work we describe the tissue distribution of fAGPrP during FIP, and its relationship with feline coronavirus (FCoV) and myeloid cells. Tissues from five control cats and from 15 cats with FIP were examined by immunohistochemistry using monoclonal antibodies against human AGP, FCoV and myeloid antigens. Diffuse fAGPrP positivity within the lesions, likely due to vascular plasma leakage, endothelial and epithelial lining were detectable. Compared to controls, fAGPrP-expressing cells often increased in number and were diffusely distributed in lymph nodes, as usually occurs for IgM-producing plasma cells during early immune responses. These findings did not depend on the presence of FCoVs or of myeloid cells, suggesting that fAGPrP is not directly involved in the pathogenesis of FIP.     

Bacillus subtilis B21 and Bacillus licheniformis B26 improve intestinal health and performance of broiler chickens with Clostridium perfringens‐induced necrotic enteritis

This study investigated the influence of Bacillus‐based probiotics on performance and intestinal health in broiler challenged with Clostridium perfringens‐induced necrotic enteritis. One‐day‐old Arbor Acre (n = 480) were randomly assigned to four treatments with 10 cages of 12 birds: (a) basal diet negative control (NC), with no probiotics nor antibiotics formulated to contain 2,930 and 3,060 kcal/kg with 24.07 and 15.98% CP, for starter and finisher diet, respectively, (b) basal diet + enramycin (5 mg/kg), an antibiotic growth promoter (AGP); (c) basal diet + Bacillus subtilis B21 at 2 × 10⁹ CFU per g (BS); (d) basal diet + Bacillus licheniformis B26 at 2 × 10⁹ CFU per g (BL); growth performance, intestinal morphology, intestinal lesion scores, short‐chain fatty acids (SCFAs) and mucosal barrier tight junction's (TJ) mRNA expression were assessed. NC‐ and BL‐fed groups showed higher (p = 0.005) average daily feed intake from d1 to d21 than AGP and BS, whereas BS‐ and AGP‐fed groups showed higher average daily weight gain from d22 to d42 and d1 to d42 of age. Higher mortality rate of (12.5%) and lower of (5.5%) were recorded in AGP and NC fed‐groups respectively, lesion score was higher in BS and BL than in AGP, while no lesion was observed in NC group, results revealed higher duodenum and jejunum villus height to crypt depth (VH:CD) compared with NC and BS. Probiotics‐fed groups showed higher total (SCFAs), acetic and butyric acid concentrations at d21 post‐challenge (PC) than other groups. The expression of claudin‐1 was upregulated in duodenum (d7) PC and in jejunum (d7) and (d21) PC in BL group, while at d21 PC, the expression of occludens was higher in jejunum and ileum by AGP and BL. The present study indicated both BS and BL have some similarity with AGP in preventing or partially preventing NE effect in broilers.