# Venomics across the Bothrops neuwiedi Species Complex Revealed a P‑III Snake Venom Metalloproteases/K49-PLA2 Dichotomy and a Remarkable Paraspecific Neutralization of the Brazilian Pentabothropic Antivenom

## TL;DR

This study analyzed snake venoms from seven species in the Bothrops neuwiedi complex, revealing toxin variability and strong antivenom effectiveness across species.

## Contribution

The study identifies a PIII-SVMP/K49-PLA2 toxin dichotomy and demonstrates the paraspecific neutralization of a Brazilian antivenom across the B. neuwiedi clade.

## Key findings

- Venoms from seven Bothrops species showed significant inter- and intraspecific variability in toxin composition and enzymatic activities.
- The Brazilian pentabothropic antivenom effectively neutralized lethality in all seven species despite venom variability.
- A PIII-SVMP/K49-PLA2 compositional dichotomy was observed, with strong immunological cross-reactivity across the Bothrops phylogeny.

## Abstract

Snakes of the Bothrops neuwiedi complex
are widely distributed and represent medically important species in
Brazil. Here, we report compositional and functional profiles of the
venom of seven species of Bothrops neuwiedi group: Bothrops mattogrossensis, Bothrops pauloensis, Bothrops pubescens, Bothrops diporus, Bothrops neuwiedi, Bothrops marmoratus, and Bothrops erythromelas. Toxin
composition of individual and pooled venoms showed remarkable inter-
and intraspecific variability of the relative abundance of toxins
(evidenced by SDS-PAGE and RP-HPLC) and enzymatic activities (proteolytic,
PLA2, and thrombin-like activities). In vivo analyses showed that B. erythromelas venom is the most hemorrhagic, B. diporus was the most lethal, and B. pubescens showed the highest myotoxic activity. Histopathological analysis
showed that all venoms induced edema, hemorrhage, inflammatory infiltrate,
and necrosis of muscle fibers. Consistent with large research evidence
on the paraspecificity of various commercial antivenoms generated
in Latin America, the pentabothropic antivenom produced by Instituto
Butantan showed a high profile of immunoreactivity and lethality neutralization
capability toward the venoms of the seven species of the B. neuwiedi clade. Interpreted through the prism
of evolution, our data revealed a PIII-SVMP/K49-PLA2 compositional
dichotomy and a remarkable conservation of immunological cross-reactivity
across congeneric venoms throughout the 12–16 million years
of Bothrops phylogeny.