Histopathology: an old yet important technique to diagnose paratuberculosis in non-descript water buffaloes

Paratuberculosis (PTB) in buffaloes is a chronic enteric disease triggering health implications and huge economic losses in livestock. This study was designed to explore a simple, cost-effective diagnostic approach for PTB in water buffaloes. Blood (5 ml/animal) and intestinal tissue samples accompanied by lymph nodes associated with mesentery were collected from weak and diarrhoeac animals slaughtered at local abattoirs. Out of total n=771 clinically suspected animals, only n=53 carcasses were sampled based on gross observation. Tissue smears of the gut mucosae were obtained and were made adopting special staining protocol. Tissue samples were processed by paraffin sectioning and stained with Ziehl-Neelsen and Hematoxylin-Eosin staining methods. Acid-fast bacilli were observed only in 11/53 cases on mucosal tissue smears. Pressure atrophy of small intestine villi were evident, and the mucosae were found sloughed off. The submucosae were heavily infiltrated with mononuclear cells and multifocal cellular nodules dominated by epithelioid macrophages. The foamy cytoplasm of the macrophages appeared to be engorged with acid fast bacilli and depicted the positive cases. All tissue sections of the suspected samples showed 100% +ve results while only 20.8% samples were found +ve with smear method. All histo-pathologically positive cases were further confirmed by ELISA based serological analysis. Therefore, it was concluded that histopathology is an economical and yet the most trusted tool for diagnosing bubalian PTB in countries like Pakistan

Exome Sequencing Revealed a Novel Splice Site Variant in the CRB2 Gene Underlying Nephrotic Syndrome

Background and Objectives: Nephrotic syndrome (NS) is a kidney disease where the patient has a classic triad of signs and symptoms including hypercholesterolemia, hypoalbuminemia, proteinuria (>3.5 g/24 h), and peripheral edema. In case of NS, the damaged nephrons (structural and functional unit of the kidney) filter unwanted blood contents to make urine. Thus, the urine contains unwanted proteins (proteinuria) and blood cells (hematuria), while the bloodstream lacks enough protein albumin (hypoalbuminemia). Nephrotic syndrome is divided into two types, primary NS, and secondary NS. Primary NS, also known as primary glomerulonephrosis, is the result of a glomerular disease that is limited to the kidney, while secondary NS is a condition that affects the kidney and other parts of the body. The main causes of primary NS are minimal change disease, membranous glomerulonephritis, and focal segmental glomerulosclerosis. In the present study we recruited a family segregating primary NS with the aim to identify the underlying genetic etiology. Such type of study is important in children because it allows counseling of other family members who may be at risk of developing NS, predicts risk of recurrent disease phenotypes after kidney transplant, and predicts response to immunosuppressive therapy. Materials and Methods: All affected individuals were clinically evaluated. Clinical examination, results of laboratory tests, and biopsy investigations led us to the diagnosis. The next-generation sequencing technique (whole-exome sequencing) followed by Sanger sequencing identified a novel homozygous splice site variant (NM_173689.7: c.941-3C>T) in the CRB2 gene. The variant was present in a homozygous state in the affected individuals, while in a heterozygous state in phenotypically normal parents. Results: The study expanded the spectrum of the mutations in the gene CRB2 responsible for causing NS. Conclusions: In addition, the study will also help in genetic counseling, carrier testing, and prenatal and/or postnatal early diagnosis of the disease in the affected family