a5b1641167d61e6bd8d9457dc2f0d769 pmc_11350667.pdf df7c0ab416f308dc7ed4d236a94d2ac10f6beae7 pmc_11350667.pdf 2945fe2cfae2b96b24465184d5b0f20e41c6223096d235450c62b62d14205742 pmc_11350667.pdf Title: Serological Investigation for Brucella ceti in Cetaceans from the Northwestern Mediterranean Sea Subject: Neurobrucellosis in cetaceans, caused by Brucella ceti, is a relevant cause of death in striped dolphins (Stenella coeruleoalba) from the Mediterranean Sea. Serological tests are not used as a routinary technique for the diagnosis of this infection. We briefly describe the pathological findings of nine free-ranging stranded cetaceans diagnosed with Brucella disease or infection in our veterinary necropsy service from 2012 to 2022. The findings included focal diskospondylitis and non-suppurative meningitis, choroiditis and radiculitis. Additionally, an exploratory serological study was conducted in sixty-six frozen sera collected in the period 2012–2022 from fifty-seven striped dolphins, five Risso’s dolphins (Grampus griseus), two common bottlenose dolphins (Tursiops truncatus), one common dolphin (Delphinus delphis) and one pilot whale (Globicephala melas) to compare antibody levels in Brucella-infected (n = 8) and non-infected (n = 58) animals, classified by the cause of death, sex, age class and cetacean morbillivirus (CeMV) infection status. The authors hypothesized that active infection in cases of neurobrucellosis would elicit a stronger, detectable humoral response compared to subclinical infections. We performed a commercial competition ELISA (cELISA) using serial serum dilutions for each sample, considering a percentage of inhibition (PI) of 40% as positive. A titer of 1:160 was arbitrarily determined as the seropositivity threshold. Seropositive species included striped dolphins and Risso’s dolphins. Seroprevalence was higher in animals with neurobrucellosis (87.5%) compared to the overall seroprevalence (31.8%) and to other causes of death, indicating, likely, a high sensitivity but low specificity for neurobrucellosis. Animals with chronic CeMV seemed to have higher seroprevalences, as well as juveniles, which also had a higher disease prevalence. These results indicate, as in other studies, that antibodies are not decisive against clinical brucellosis, although they may indicate a carrier state, and that CeMV may influence Brucella epidemiology. More research is required to elucidate the epidemiology and pathogenesis and to resolve the complicated host–pathogen interaction in Brucella species. Keywords: brucellosis; serology; antibodies; ELISA; morbillivirus; dolphin; stranding; necropsy Author: Laura Martino, María Cuvertoret-Sanz, Sarah Wilkinson, Alberto Allepuz, Albert Perlas, Llilianne Ganges, Lola Pérez and Mariano Domingo Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Fri Aug 23 09:07:27 2024 CEST ModDate: Fri Aug 23 09:14:40 2024 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 13 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2725384 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- YSPQJI+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 IDTDJP+URWPalladioL-Bold Type 1 Custom yes yes yes 16 0 ZXEBEH+URWPalladioL-Roma Type 1 Custom yes yes yes 22 0 TEAOWH+URWPalladioL-Ital Type 1 Custom yes yes yes 27 0 XEINJN+URWPalladioL-BoldItal Type 1 Custom yes yes yes 32 0 OHXQCJ+CMSY10 Type 1 Builtin yes yes yes 37 0 UOHDZX+VnURWPalladioL Type 1 Custom yes yes yes 42 0 SZMRXJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 58 0 OLIAAM+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 86 0 OLIABM+PalatinoLinotype CID TrueType Identity-H yes yes yes 89 0 OLIABN+PalatinoLinotype TrueType WinAnsi yes yes no 95 0 OLIABP+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 98 0 OLIAGA+PalatinoLinotype,BoldItalic TrueType WinAnsi yes yes no 101 0 OLIAHB+PalatinoLinotype TrueType MacRoman yes yes no 104 0 OLIAAM+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 132 0 OLIABM+PalatinoLinotype CID TrueType Identity-H yes yes yes 135 0 OLIABN+PalatinoLinotype TrueType WinAnsi yes yes no 141 0 OLIABP+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 144 0 OLIAGA+PalatinoLinotype,BoldItalic TrueType WinAnsi yes yes no 147 0 OLIAHB+PalatinoLinotype TrueType MacRoman yes yes no 150 0 OLIAAM+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 170 0 OLIABM+PalatinoLinotype CID TrueType Identity-H yes yes yes 173 0 OLIABN+PalatinoLinotype TrueType WinAnsi yes yes no 179 0 OLIABP+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 182 0 OLIAAM+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 196 0 OLIABM+PalatinoLinotype CID TrueType Identity-H yes yes yes 199 0 OLIABN+PalatinoLinotype TrueType WinAnsi yes yes no 205 0 OLIABP+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 208 0 OLIAHB+PalatinoLinotype TrueType MacRoman yes yes no 211 0 JQNTEN+MSAM10 Type 1 Builtin yes yes yes 220 0 CKBPVA+EURM10 Type 1 Builtin yes yes yes 231 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-09-19 02:49:28 CEST RepresentationInformation: pmc_11350667.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-09-18 02:31:09 CEST Size: 2725384 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 420 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Cases Investigated and Necropsy Procedure Destination: subsection.2.1 Item: Title: Serology for Brucella Destination: subsection.2.2 Item: Title: Brucella Isolation Destination: subsection.2.3 Item: Title: Age Determination Destination: subsection.2.4 Item: Title: Data Analysis Destination: subsection.2.5 Item: Title: Results Destination: section.3 Children: Item: Title: Pathological Investigation Destination: subsection.3.1 Item: Title: Culture of Brucella Destination: subsection.3.2 Item: Title: Serological Investigation Destination: subsection.3.3 Children: Item: Title: Serological Results Compared by Cause of Death Destination: subsubsection.3.3.1 Item: Title: Serology by Age Class Destination: subsubsection.3.3.2 Item: Title: Brucella Serology in CeMV-Infected Cetaceans Destination: subsubsection.3.3.3 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: appendix.A. Info: Title: Serological Investigation for Brucella ceti in Cetaceans from the Northwestern Mediterranean Sea Author: Laura Martino, María Cuvertoret-Sanz, Sarah Wilkinson, Alberto Allepuz, Albert Perlas, Llilianne Ganges, Lola Pérez and Mariano Domingo Subject: Neurobrucellosis in cetaceans, caused by Brucella ceti, is a relevant cause of death in striped dolphins (Stenella coeruleoalba) from the Mediterranean Sea. Serological tests are not used as a routinary technique for the diagnosis of this infection. We briefly describe the pathological findings of nine free-ranging stranded cetaceans diagnosed with Brucella disease or infection in our veterinary necropsy service from 2012 to 2022. The findings included focal diskospondylitis and non-suppurative meningitis, choroiditis and radiculitis. Additionally, an exploratory serological study was conducted in sixty-six frozen sera collected in the period 2012 2022 from fifty-seven striped dolphins, five Risso’s dolphins (Grampus griseus), two common bottlenose dolphins (Tursiops truncatus), one common dolphin (Delphinus delphis) and one pilot whale (Globicephala melas) to compare antibody levels in Brucella-infected (n = 8) and non-infected (n = 58) animals, classified by the cause of death, sex, age class and cetacean morbillivirus (CeMV) infection status. The authors hypothesized that active infection in cases of neurobrucellosis would elicit a stronger, detectable humoral response compared to subclinical infections. We performed a commercial competition ELISA (cELISA) using serial serum dilutions for each sample, considering a percentage of inhibition (PI) of 40% as positive. A titer of 1:160 was arbitrarily determined as the seropositivity threshold. Seropositive species included striped dolphins and Risso’s dolphins. Seroprevalence was higher in animals with neurobrucellosis (87.5%) compared to the overall seroprevalence (31.8%) and to other causes of death, indicating, likely, a high sensitivity but low specificity for neurobrucellosis. Animals with chronic CeMV seemed to have higher seroprevalences, as well as juveniles, which also had a higher disease prevalence. These results indicate, as in other studies, that antibodies are not decisive against clinical brucellosis, although they may indicate a carrier state, and that CeMV may influence Brucella epidemiology. More research is required to elucidate the epidemiology and pathogenesis and to resolve the complicated host pathogen interaction in Brucella species. 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